Benzyl morpholine derivatives

ABSTRACT

A compound of formula (I) (I) wherein R is H; Ar is an aromatic group selected from phenyl; X is a phenyl group; R′ is H or C1-C4 alkyl; each R1 is independently H or C1-C4 alkyl; and pharmaceutically acceptable salts thereof

This is the national phase application, under 35 USC 371, forPCT/US2003/023268, filed 1 Aug. 2003, which claims the benefit, under 35USC 119(e), of U.S. provisional application No. 60/415,327, filed 1 Oct.2002 and, under 35 USC 119(a), of GB application 0219685.5, filed 23Aug. 2002.

This invention relates to novel benzyl morpholine compounds, and totheir use in inhibiting serotonin and norepinephrine reuptake.

Serotonin has been implicated in the aetiology of many disease statesand has been found to be of importance in mental illnesses, depression,anxiety, schizophrenia, eating disorders, obsessive compulsive disorder(OCD) and migraine. Indeed many currently used treatments of thesedisorders are thought to act by modulating serotonergic tone. During thelast decade, multiple serotonin receptor subtypes have beencharacterised. This has led to the realisation that many treatments actvia the serotonergic system, such as selective serotonin reuptakeinhibitor (SSRI) antidepressants which increase serotonin transmission,for example, the hydrochloride salt of fluoxetine.

Drugs that exert their main action on the norepinephrinergic system havebeen available for some time, however their lack of selectivity has madeit difficult to determine specific clinical effects produced by aselective action on norepinephrine reuptake. Accumulating evidenceindicates that the norepinephrinergic system modulates drive and energy,whereas the serotonergic system modulates mood. Thus norepinephrineappears to play an important role in the disturbances of vegetativefunction associated with affective, anxiety and cognitive disorders.Atomoxetine hydrochloride is a selective inhibitor of norepinephrinereuptake, and is marketed for the treatment of attention deficithyperactivity disorder (ADHD). Reboxetine is also a selectivenorepinephrine reuptake inhibitor and is marketed for the treatment ofdepression. WO99/15177 discloses the use of Reboxetine to treat ADHD andWO01/01973 discloses the use of S,S-Reboxetine to treat ADHD.

Norepinephrine and serotonin receptors are known to interactanatomically and pharmacologically. Compounds that affect only serotoninhave been shown to exhibit modulatory effects on norepinephrine,pointing toward an important relationship between the twoneurotransmitter systems.

Duloxetine, (+)-N-methyl-3-(1-naphthalenyloxy)-2-thiophenepropanaminehydrochloride, inhibits the reuptake of both norepinephrine andserotonin, and is currently under development for the treatment ofdepression and urinary incontinence. The compound duloxetine wasdisclosed in U.S. Pat. Nos. 5,023,269 and 4,956,388.

According to the present invention there is provided a compound offormula (I)

wherein

-   R is H;-   Ar is an aromatic group selected from phenyl; X is a phenyl group;    R′ is H or C₁-C₄ alkyl; and each R₁ is independently H or C₁-C₄    alkyl; and pharmaceutically acceptable salts thereof.

The aromatic group Ar may be substituted or unsubstituted phenyl. Forexample, Ar may be unsubstituted phenyl or, preferably, phenylsubstituted with 1, 2, 3, 4 or 5 substitutents, preferably with 1 or 2,for example 1, substituent. The substituted phenyl group is preferablysubstituted in the 2-position. Suitable substitutents include C₁-C₄alkyl, O(C₁-C₄ alkyl), S(C₁-C₄ alkyl), halo, and phenyl, optionallysubstituted with, for example, halo, C₁-C₄ alkyl or O(C₁-C₄ alkyl).

The group X may be substituted or unsubstituted phenyl. For example, Xmay be phenyl substituted with 1, 2, 3, 4 or 5 substituents, preferablywith 1 substituent. Suitable substituents include C₁-C₄ alkyl, O(C₁-C₄alkyl), and halo.

“C₁-C₄ alkyl” as used herein includes straight and branched chain alkylgroups of 1, 2, 3 or 4 carbon atoms, and may be unsubstituted orsubstituted. C₁-C₂ alkyl groups are preferred. Suitable substituentsinclude halo. Thus the term “C₁-C₄ alkyl” includes haloalkyl. Similarterms defining different numbers of C atoms (e.g. “C₁-C₂ alkyl”) take ananalogous meaning. When R′ is C₁-C₄ alkyl it is preferablyunsubstituted. When R¹ is C₁-C₄ alkyl it is preferably unsubstituted.

“Halo” includes F, Cl, Br and I, and is preferably F or Cl.

For the compounds of formula (I) above, R′ is preferably H or Me. Morepreferably R′ is H.

For the compounds of formula (I) above, each R¹ is preferably H or Mewith 0, 1, 2 or 3 of R¹ being Me. More preferably only 1 R¹ is Me. Mostpreferably all R¹ are H.

For the compounds of formula (I) above, it is preferred that R′ and allR¹ are H.

Particularly preferred substituents for the Ar group includetrifluoromethyl and methoxy.

A preferred group of compounds according to the present invention isrepresented by formula (II);

in which R₂ and R₃ are each independently selected from H, C₁-C₄ alkyl,O(C₁-C₄ alkyl), S(C₁-C₄ alkyl), halo, and phenyl; and

-   R₄ is selected from H, C₁-C₄ alkyl, and O(C₁-C₄ alkyl) and halo; and    pharmaceutically acceptable salts thereof.

R₂ is preferably C₁-C₂ alkyl, O(C₁-C₂ alkyl), S(C₁-C₂ alkyl), Cl or F.R₃ is preferably H, Me or Cl. R₄ is preferably H, C₁-C₂ alkyl, O(C₁-C₂alkyl), Cl or F.

The compounds of the present invention are dual reuptake inhibitors ofserotonin and norepinephrine. Biogenic amine transporters control theamount of biogenic amine neurotransmitters in the synaptic cleft.Inhibition of the respective transporter leads to a rise in theconcentration of that neurotransmitter within the synaptic cleft.Compounds of Formula (I) and their pharmaceutically acceptable saltspreferably exhibit a K_(i) value less than 100 nM at the norepinephrinetransporter and a K_(i) value less than 100 nM at the serotonintransporter as determined using the scintillation proximity assays asdescribed below. More preferred compounds of Formula (I) and theirpharmaceutically acceptable salts exhibit a K_(i) value less than 50 nMat the norepinephrine transporter and a K_(i) value less than 50 nM atthe serotonin transporter. Especially preferred compounds of Formula (I)and their pharmaceutically acceptable salts exhibit a K_(i) value lessthan 20 nM at the norepinephrine transporter and a K_(i) value less than20 nM at the serotonin transporter. Preferably, compounds of the presentinvention selectively inhibit the norepinephrine and serotonintransporters relative to the dopamine transporter by a factor of atleast five, more preferably by a factor of at least ten. Advantageously,they have a reduced interaction (both as substrate and inhibitor) withthe liver enzyme Cytochrome P450 (CYP2D6). That is to say, theypreferably exhibit less than 75% metabolism via the CYP2D6 pathwayaccording to the CYP2D6 substrate assay described below and theypreferably exhibit an IC50 of >6 μM according to the CYP2D6 inhibitorassay described below. They are particularly useful for the treatment ofdisorders associated with serotonin and norepinephrine dysfunction inmammals, such as CNS disorders including depression, persistant pain andstress urinary incontinence.

The term “serotonin and norepinephrine dysfunction” as used hereinrefers to a reduction in the amount of serotonin and norepinephrineneurotransmitter within the synaptic cleft below that which would beconsidered to be normal. Thus the phrase “disorders associated withserotonin and norepinephrine dysfunction in mammals” refers to disorderswhich are associated with a reduction in the amount of serotonin andnorepinephrine neurotransmitter within the synaptic cleft below thatwhich would be considered to be normal for the mammalian species inquestion. Some examples of disorders currently believed to be associatedwith reduced levels of serotonin and norepinephrine within the synapticcleft are detailed below.

The compounds of the present invention are also indicated for thetreatment of disorders which are ameliorated by an increase in theamount of serotonin and norepinephrine neurotransmitter within thesynaptic cleft of a mammal above that which would be considered to benormal for the mammalian species in question.

The term “treatment” as used herein refers to both curative andprophylactic treatment of disorders associated with serotonin andnorepinephrine dysfunction.

Compounds of the present invention may be prepared by reacting acompound of the formula (III):

where R5 is a protecting group, e.g. benzyl, X, R′ and R¹ are as formulaI above and Y is a leaving group, with an aryl thiol. Examples ofsuitable leaving groups include halo and mesylate, but the nature of theleaving group is not critical.

Compounds of the present invention may also be prepared by deprotectinga compound of the formula (IV):

where R₅ is a protecting group, e.g. benzyl, and Ar, X, R′ and R¹ are asdefined in formula (I) above to provide a compound of formula (I),optionally followed by the step of forming a pharmaceutically acceptablesalt.

Suitable N-protecting groups will be known to the person skilled in theart as will methods for their removal. Further information on suitabledeprotecting groups is contained in the well known text “ProtectiveGroups in Organic Synthesis”, Theodora W. Greene and Peter G. M. Wuts,John Wiley & Sons, Inc., New York, 1999, pp. 494-653. PreferredN-protecting groups include benzyl, allyl, carbamates such asbenzyloxycarbonyl (cbz) and t-butyloxycarbonyl (boc) and amides.

Compounds of the present invention may be prepared by conventionalorganic chemistry techniques from N-benzyl-cyanomorpholine 1 (Route A)or N-benzyl-morpholinone 2 (Route B) as outlined in Scheme 1 below. Forclarity, X is shown as phenyl and R′ and R¹ are shown as H. It will beappreciated that analogous methods could be applied for other possibleidentities of X, R′ and R¹.

More detail of Route A is given in Scheme 2:

The amino alcohol can be obtained by reaction ofN-benzyl-cyanomorpholine with a Grignard reagent, followed by acidhydrolysis to give racemic phenyl ketone which may be separated onchiral HPLC. (2R)-Phenyl ketone may then be reduced with DIP-Cl to givethe amino alcohol in high diastereomeric excess. The amino alcohol maybe converted into the benzyl bromide to give the desired N-substitutedaryl thio morpholines after displacement with the requisite aryl thiol.Deprotection of the tertiary amine gives the final products.Deprotection may be achieved by reaction with α-chloroethylchloroformate as described in the specific examples. Deprotection mayalso be achieved by reaction with phenylchloroformate followed byquenching of the generated benzyl chloride with dimethylamine prior toisolation of the phenylcarbamate intermediate which is then hydrolysedin isopropanol in the presence of 30% NaOH.

Detail of Route B is Given in Scheme 3:

Treatment of N-benzyl morpholinone with a strong base such as lithiumdiisopropylamide at low temperature followed by addition of benzaldehydegives aldol adducts as a 2:1 mixture of diastereomer pairs, which may beseparated using conventional chromatographic techniques. Reduction witha borane reagent at elevated temperatures gives diasteremeric aminoalcohol pairs.

Amino alcohol pair (2S,2'S) and (2R,2'R) may be converted to bromide andfurther to racemic aryl thio morpholines as outlined in Scheme 4. Aminoalcohol pair (2R,2'S) and (2S,2'R) may be converted into thecorresponding mesylate. Displacement with the requisite thiol, followedby removal of the nitrogen protecting group furnishes aryl thiolmorpholines as racemic mixtures of two diastereomers. The racemic arylthiol morpholines may be separated into enantiomerically pure productsusing chiral HPLC technology.

Compounds of the present invention are selective inhibitors of thereuptake of both serotonin and norepinephrine and as such are useful aspharmaceuticals. They may be indicated in the treatment of disordersassociated with serotonin and norepinephrine dysfunction in mammals,including depression, eating disorders (including bulimia and anorexianervosa), inflammatory bowel disorders, functional bowel disorders,dyspepsia, chron's disease, iletis, ischemic bowel disease, ulcerativecolitis, gastroesophageal reflux for functional bowel disorders,irritable bowel syndrome, obesity, insterstitial cystitis, urethralsyndrome, gastric motility disorders, substance abuse (includingalcoholism, tobacco abuse, symptoms caused by withdrawal or partialwithdrawal from the use of tobacco or nicotine and drug addictionincluding cocaine abuse), smoking cessation, pain (includinginflammatory pain, neuropathic pain, non-neuropathic non-inflammatorypain, persistent pain, persistent pain of inflammatory and/orneuropathic origin, headache and migraine), urinary incontinence(including stress urinary incontinence and urge incontinence), dementiaof ageing, senile dementia, Alzheimer's, memory loss, Parkinsonism,attention-deficit disorder (including attention-deficit hyperactivitydisorder), anxiety, social phobia, disruptive behavior disorders,conduct disorders, impulsive control disorders, borderline personalitydisorder, chronic fatigue syndrome, panic disorders, obsessivecompulsive disorder, post-traumatic stress disorder, schizophrenia,gastrointestinal disorders, cardiovascular disorders, emesis, sleepdisorders, cognitive disorders, psychotic disorders, brain trauma,premenstrual syndrome or late luteal syndrome, sexual dysfunction(including premature ejaculation and erectile difficulty), autism,mutism and trichotilomania. The compounds of the present invention areparticularly suitable for the treatment of pain.

For clinical purposes, pain may be divided into two categories: acutepain and persistent pain. Acute pain is provoked by noxious stimulationproduced by injury and/or disease of skin, deep somatic structures orviscera, or abnormal function of muscle or viscera that does not produceactual tissue damage. On the other hand, persistent pain can be definedas pain that persists beyond the usual course of an acute disease or areasonable time for an injury to heal or that is associated with achronic pathologic process that causes continuous pain or the painrecurs at intervals for months or years. If pain is still present aftera cure should have been achieved, it is considered persistent pain. Forthe purpose of the present invention, persistent pain can be chronicnon-remitting or recurrent. The difference in definition between acuteand persistent pain is not merely semantic but has an important clinicalrelevance. For example, a simple fracture of the wrist usually remainspainful for a week to 10 days. If the pain is still present beyond thetypical course of treatment, it is likely that the patient is developingreflex sympathetic dystrophy, a persistent pain syndrome that requiresimmediate effective therapy. Early and effective interventionpotentially prevents the undue disability and suffering, and avoids thepotential development of a condition that becomes refractory to therapy.

Acute and chronic pain differ in etiology, mechanisms, pathophysiology,symptomatology, diagnosis, therapy, and physiological responses. Incontrast to the transitory nature of acute pain, persistent pain iscaused by chronic pathologic processes in somatic structures or viscera,by prolonged and sometimes permanent dysfunction of the peripheral orcentral nervous system, or both. Also, persistent pain can sometimes beattributed to psychologic mechanisms and/or environmental factors.

Current therapies for persistent pain include opiates, barbiturate-likedrugs such as thiopental sodium and surgical procedures such asneurectomy, rhizotomy, cordotomy, and cordectomy.

References herein to pain are intended to refer to persistent pain.

The present invention provides pharmaceutical compositions comprising acompound of formula I or formula II or a pharmaceutically acceptablesalt thereof, together with a pharmaceutically acceptable diluent orcarrier.

Further, the present invention provides a compound of formula I or apharmaceutically acceptable salt thereof, for use as a pharmaceutical;and a compound of formula I or a pharmaceutically acceptable saltthereof, for use as a selective inhibitor of the reuptake of bothserotonin and norepinephrine. Preferably such selective inhibitionoccurs within mammalian cells (including mammalian cell membranepreparations), especially those found within the central and/orperipheral nervous system. More preferably such selective inhibitionoccurs within the cells of the central nervous system of a mammal,especially a human, in need thereof.

The present invention also provides the use of a compound of formula Ior formula II, or a pharmaceutically acceptable salt thereof, in themanufacture of a medicament for selectively inhibiting the reuptake ofserotonin and norepinephrine; the use of a compound of formula I orformula II, or a pharmaceutically acceptable salt thereof, in themanufacture of a medicament for the treatment of disorders associatedwith serotonin and norepinephrine dysfunction in mammals; the use of acompound of formula I or formula II, or a pharmaceutically acceptablesalt thereof, in the manufacture of a medicament for the treatment of adisorder selected from depression, OCD, anxiety, memory loss, urinaryincontinence, conduct disorders, ADHD, obesity, alcoholism, smokingcessation and pain; and the use of a compound of formula I or formulaII, or a pharmaceutically acceptable salt thereof, in the manufacture ofa medicament for the treatment of a disorder selected from depression,stress urinary incontinence, and persistent pain. The present inventionfurther provides a compound of formula I or formula II for treatingdisorders associated with serotonin and norepinephrine dysfunction inmammals, for example a disorder selected from depression, OCD, anxiety,memory loss, urinary incontinence, conduct disorders, ADHD, obesity,alcoholism, smoking cessation and pain, especially depression, stressurinary incontinence, and persistent pain.

Further the present invention provides a method for selectivelyinhibiting the reuptake of serotonin and norepinephrine in mammals,comprising administering to a patient in need thereof an effectiveamount of a compound of formula I or formula II or a pharmaceuticallyacceptable salt thereof; a method for treating disorders associated withserotonin and norepinephrine dysfunction in mammals, comprisingadministering to a patient in need thereof an effective amount of acompound of formula I or formula II or a pharmaceutically acceptablesalt thereof; and a method for treating a disorder selected fromdepression, OCD, anxiety, memory loss, urinary incontinence, conductdisorders, ADHD, obesity, alcoholism, smoking cessation and pain,especially depression, stress urinary incontinence or persistant pain,comprising administering to a patient in need thereof an effectiveamount of a compound of formula I or a pharmaceutically acceptable saltthereof.

The present invention includes the pharmaceutically acceptable salts ofthe compounds of formula I and formula II. Suitable salts include acidaddition salts, including salts formed with inorganic acids, for examplehydrochloric, hydrobromic, nitric, sulphuric or phosphoric acids, orwith organic acids, such as organic carboxylic or organic sulphonicacids, for example, acetoxybenzoic, citric, glycolic, o-mandelic-1,mandelic-dl, mandelic d, maleic, mesotartaric monohydrate,hydroxymaleic, fumaric, lactobionic, malic, methanesulphonic, napsylic,naphthalenedisulfonic, naphtoic, oxalic, palmitic, phenylacetic,propionic, pyridyl hydroxy pyruvic, salicylic, stearic, succinic,sulfanilic, tartaric, 2-hydroxyethane sulphonic, toluene-p-sulphonic,and xinafoic acids.

In addition to the pharmaceutically acceptable salts, other salts areincluded in the invention. They may serve as intermediates in thepurification of compounds or in the preparation of other, for examplepharmaceutically acceptable, acid addition salts, or are useful foridentification, characterisation or purification.

It will be appreciated that compounds of formula I and formula IIpossess one or more asymmetric carbon atoms, and that the presentinvention is directed specifically to individual stereoisomers. Theparticular stereochemistry of the present compounds is essential to thepharmacological profile of the compounds. In the present specification,where a structural formula does not specify the stereochemistry at oneor more chiral centres, it encompasses all possible stereoisomers andall possible mixtures of stereoisomers (including, but not limited to,racemic mixtures), which may result from stereoisomerism at each of theone or more chiral centers.

The compounds of the present invention may be used as medicaments inhuman or veterinary medicine. The compounds may be administered byvarious routes, for example, by oral or rectal routes, topically orparenterally, for example by injection, and are usually employed in theform of a pharmaceutical composition.

Such compositions may be prepared by methods well known in thepharmaceutical art and normally comprise at least one active compound inassociation with a pharmaceutically acceptable diluent, excipient orcarrier. In making the compositions of the present invention, the activeingredient will usually be mixed with a carrier or diluted by a carrier,and/or enclosed within a carrier which may, for example, be in the formof a capsule, sachet, paper or other container. Where the carrier servesas a diluent, it may be solid, semi-solid, or liquid material which actsas a vehicle, excipient or medium for the active ingredient. Thus, thecomposition may be in the form of tablets, lozenges, sachets, cachets,elixirs, suspensions, solutions, syrups, aerosols (as a solid or in aliquid medium), ointments containing, for example, up to 10% by weightof the active compound, soft and hard gelatin capsules, suppositories,injection solutions and suspensions and sterile packaged powders.

Some examples of suitable carriers are lactose, dextrose, vegetableoils, benzyl alcohols, alkylene glycols, polyethylene glycols, glyceroltriacetate, gelatin, carbohydrates such as starch and petroleum jelly,sucrose sorbitol, mannitol, starches, gum acacia, calcium phosphate,alginates, tragacanth, gelatin, syrup, methyl cellulose, methyl- andpropyl- hydrobenzoate, talc, magnesium stearate and mineral oil. Thecompounds of formula (I) can also be lyophilized and the lyophilizatesobtained used, for example, for the production of injectionpreparations. The preparations indicated can be sterilized and/or cancontain auxiliaries such as lubricants, preservatives, stabilizersand/or wetting agents, emulsifiers, salts for affecting the osmoticpressure, buffer substances, colourants, flavourings and/or one or morefurther active compounds, e.g. one or more vitamins. Compositions of theinvention may be formulated so as to provide, quick, sustained ordelayed release of the active ingredient after administration to thepatient by employing procedures well known in the art.

The compositions are preferably formulated in a unit dosage form, eachdosage unit containing from about 5 to about 500 mg, more usually about25 to about 300 mg, of the active ingredient. The term “unit dosageform” refers to physically discrete units suitable as unitary doses forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalcarrier.

The following Examples illustrate compounds of the present invention andmethods for their synthesis.

Stereochemical Conventions

The absolute stereochemistry of compounds according to the presentinvention may be determined by reference to X-ray crystallography forthe following (2S,2′S) compound

X-ray crystallographic data for the above compound is listed in Tables1-6 herein.

All of the Examples herein were obtained as single isomers eitherthrough the use of chirally pure starting material or chiral separationmethods, such as HPLC.

EXAMPLE 1(2R)-2-((R)-[4-methoxyphenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

i) (+/−)-[4-Methoxyphenyl][(4-benzylmorpholin-2-yl]methanone

To stirred magnesium turnings (5.4 g, 0.22 mol) in dry THF (20 ml) atroom temperature under nitrogen was added sufficient 1,2-dibromoethane(ca. 0.3 ml) to create an exotherm. A solution of 4-bromoanisole (13.90g, 74.25 mmol) in dry THF (25 ml) was then added dropwise at a rate tomaintain a gentle reflux. After addition, allowed to cool to below 30°C. then added over 5 min. period to a stirred solution of4-benzyl-2-cyanomorpholine (5.0 g, 24.75 mmol) in dry THF (50 ml) cooledto −20° C. under nitrogen. After addition stirred at room temperaturefor 30 min. then cooled to 0° C. and added 5M HCl (25 ml) dropwise.After 5 min. stirring, made basic by addition of 2M NaOH and theresulting suspension filtered through celite. The aqueous phase wasseparated and washed with diethyl ether (2×). The combined organicphases was dried over magnesium sulphate, filtered and evaporated to ayellow oil. The oil was purified by flash chromatography on silicaeluting with ethyl acetate/heptane gradient 25/75 to 70/30 to giverequired product as a yellow oil (5.46 g).

ii) (R)-[4-methoxyphenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(S)-[4-methoxyphenyl][(2R)-4-benzylmorpholin-2-yl]methanol

To a stirred solution of(+/−)-[4-Methoxyphenyl][4-benzylmorpholin-2-yl]methanone (5.40 g, 17.36mmol) in methanol (60 ml) at 5° C. was added sodium borohydride (1.31 g,34.72 mmol) portionwise. The mixture was stirred at room temperature for1.5 h, cooled to 10° C. and added water to terminate reaction.Concentrated in vacuo, diluted with water and extracted with ethylacetate (2×). Extracts washed with water and brine, dried over magnesiumsulphate, filtered and evaporated to an oil. The crude mixture ofdiastereomers was purified and separated by flash chromatography onsilica eluting with diethyl ether/toluene (3/2) to give the titlediastereomer as a colourless oil (2.14 g).

iii)(2R)-2-((R)-[4-methoxyphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylymorpholineand(2S)-2-((S)-[4-metloxyphenyl]{[2-nethoxyphenyl]thio}methyl)-4-benzylmorpholine

A mixture of (R)-[4-methoxyphenyl][(2S)-4-benzylmorpholin-2-yl]methanoland (S)-[4-methoxyphenyl][(2R)-4-benzylmorpholin-2-yl]methanol (118 mg,0.376 mmol), 2,2′-dimethoxydiphenyldisulphide (210 mg, 0.75 mmol) andtributylphosphine (152 mg, 1.50 mmol) in dry THF (2 ml) was heated atreflux under nitrogen overnight. The reaction mixture was cooled to roomtemperature and evaporated to an oil. The crude oil was purified byflash chromatography on silica eluting with heptane/ethyl acetate(4/1then 3/2) to give the product as a colourless oil (95 mg).

iv)(2R)-2-((R)-[4-methoxyphenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

Stirred (2S)-2-((S)-[4-methoxyphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine(2R)-2-((R)-[4-methoxyphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine (618 mg, 1.42 mmol) with solid supportedHunig's base (2.40 g, 8.52 mmol) and α-chloroethyl chloroformate (2.0 g,14.21 mmol) in dichloromethane (12 ml) at room temperature undernitrogen for 4 h. Filtered and concentrated in vacuo, dissolved oil inmethanol and heated at 60° C. for 1.5 h. Cooled to room temperature andpurified by SCX column chromatography eluting with ammonia/methanol (ca.3M) gave a colourless oil. The desired diastereomer was separated onchiralcel-OJ column eluting with heptane/ethanol/dimethylethylamine(20/80/0.2): 8.98 min. The required product was then obtained (60% de)after chiral chromatography on chiralpak-OD column eluting withheptane/isopropanol (70/30): 17.41 min. It was converted into the HClsalt, NMR (DMSO) 9.39 (2H, br. s), 7.3-7.1 (4H, m), 6.94-6.72 (4H, m),4.6-4.5 (1H, m), 4.12-3.92 (2H, m), 3.85-3.62 (7H, m), 3.46-3.32 (1H,m), 3.20-3.08 (1H, m), 3.04-2.89 (2H, m). LCMS: m/z 346 [M+H]⁺ @ Rt 4.24min.

EXAMPLE 2(2R)-2-((R)-(2-Fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

i) (2R)-2-[(R)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one,(2S)-2-[(S)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one,(2R)-2-[(S)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one

To a stirred solution of (+/−)-4-benzylmorpholin-3-one (10.0 g, 0.052mol) and 2-fluorobenzaldehyde (7.74 g, 0.062 mol) in dry THF (80 ml)cooled under nitrogen to −78° C. was added dropwise a solution oflithium diisopropylamide in heptane/THF/ethylbenzene (2M, 31.2 ml).After addition, stirred at −78° C. for 0.5 h then allowed to warm to 0°C. before quenching with aqueous saturated ammonium chloride.Concentrated in vacuo and extracted with dichloromethane (2×). Theextracts were dried over magnesium sulphate, filtered and evaporated toan oil. Purified on a pad of flash silica eluting with heptane/ethylacetate (100/0, 80/20, 60/40 and 50/50) to give a 1:1 mixture of thediastereomers as a colourless oil (12.05 g).

ii) (R)-[2-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(S)-[2-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol

To a stirred solution of(2R)-2-[(R)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one,(2S)-2-[(S)-(2-fluorophenyl)(hydroxy)methyl]4-benzylmorpholin-3-one,(2R)-2-[(S)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(2-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(12.0 g, 0.038 mol) in dry THF (80 ml) under nitrogen at roomtemperature was added a solution of borane in THF (1M, 150 ml). Thesolution was heated at 60° C. for ca. 4 h then at room temperatureovernight. Cooled solution to 0° C. and added dropwise methanol (68 ml)followed by 1NHCl (68 ml). The resulting mixture was heated at 60° C.for 1 h, cooled and concentrated in vacuo. The precipitate was removedby filtration and the filtrate made basic with aqueous sodium carbonate.Extracted with diethyl ether (3×), extracts washed with water and brine,dried over magnesium sulphate, filtered and evaporated to an oil. Thecrude oil was purified and partially separated by flash chromatographyon silica eluting with heptane/ethyl acetate (40/60 to 25/75) to givethe product as a colourless oil (0.713 g).iii) (R)-[2-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methylmethanesulphonate and(S)-[2-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate

To a stirred solution of(R)-[2-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(S)-[2-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol (465 mg, 1.54mmol) in dry dichloromethane (10 ml) at room temperature under nitrogenwas added triethylamine (202 mg, 2.0 mmol) and methanesulphonyl chloride(177 mg, 1.54 mmol). After 15 h, evaporated to an oil and purified byflash chromatography on silica eluting with ethyl acetate/heptane (1/1)to give the product mesylate as a colourless oil (445 mg).

iv)(2R)-2-((R)-[2-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[2-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine

To a stirred suspension of(R)-[2-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methylmethanesulphonate and(S)-[2-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate (445 mg, 1.17 mmol) and anhydrous potassium carbonate(0.97 g, 7.02 mmol) in dry degassed DMF (8 ml) under nitrogen at roomtemperature was added 2-methoxybenzenethiol (0.82 g, 5.87 mmol). Afterstirring at room temperature for 18 h, diluted with water and extractedwith diethyl ether (2×). The extracts were washed with 2NaOH, water andbrine, dried over magnesium sulphate, filtered and evaporated to an oil.

After purification by flash column chromatography (eluent:heptane/ethylacetate 80/20 [v/v]) the title product was obtained as a colourless oil(357 mg); MW 423.55; C₂₅H₂₆FNO₂S; ¹H NMR (CDCl₃): 6.65-7.5 (13H, m), 4.9(1H, d, 7 Hz), 3.9-4.05 (2H, m), 3.8 (3H, s), 3.6 (1H, dt, 8 Hz and 1Hz), 3.45 (1H, d, 13.1 Hz), 3.15 (1H, d, 13.1 Hz), 2.60 (2H, t, 8 Hz),2.05-2.2 (2H, m); FIA: m/z 424 [M+H]⁺.

v) (2R)-2-((R)-(2-Fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

Reaction of the mixture of(2R)-2-((R)-[2-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[2-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine(430 mg, 1.02 mmol) following procedure described in EXAMPLE 1(iv) gavea colourless oil (340 mg, 90% yield) from which the first elutingenantiomer was obtained after chiral chromatography on a Chiralcel-ODcolumn eluant heptane/ethanol/dimethylethylamine (40/60/0.2): Rt 10.41min. LC purity=98.6 (UV_(254nm)); MW 333.43; C₁₈H₂₀FNOS.; FIA: m/z 334[M+H]⁺. This was converted into the hydrochloride salt. ¹H NMR (CDCl₃)freebase: 7.2-7.3 (1H, m), 6.85-7.2 (8H, m), 4.85 (1H, d, 8 Hz),3.95-4.15 (2H, m), 3.85-3.9 (3H, m), 3.7 (1H, dt, 1 Hz and 7 Hz),2.6-3.0 (4H, m).

EXAMPLE 3 (2R)-2-((R)-[2,5-dichlorophenyl][phenylthio]methyl)morpholinehydrochloride

i)(2R)-2-((R)-[2,5-dichlorophenyl][phenylthio]methyl)-4-benzylmorpholine

Reacted (2R)-2-[(S)-bromo(phenyl)methyl]-4-benzylmorpholine (150 mg,0.43 mmol) (see example 8(v) method 2), 2,5-dichlorobenzenethiol (233mg, 1.30 mmol) and anhydrous potassium carbonate (71 mg, 0.52 mmol)following example 2(iv). The reaction mixture was purified directly bySCX chromatography eluting with ammonia/methanol (ca.3M) to give theproduct as an oil (174 mg).

ii) (2R)-2-((R)-[2,5-dichlorophenyl][phenylthio]methyl)morpholinehydrochloride

Debenzylation of(2R)-2-((R)-[2,5-dichlorophenyl][phenylthio]methyl)-4-benzylmorpholine(174 mg, 0.39 mmol) with polymer supported Hunig's base (0.20 g, 0.78mmol) and α-chloroethyl chloroformate (111 mg, 0.78 mmol) following theprocedure described in example 1(iv) gave after SCX chromatography theproduct as an oil (136 mg).

NMR (CDCl₃) 7.31-7.14 (7H, m), 7.00 (1H, d), 4.41 (1H, d), 4.06-3.98(1H, m), 3.90-3.82 (1H, m), 3.7-3.6 (1H, m), 2.94-2.76 (2H, m), 2.65(2H, d). m/z [M+H] 354/6/8. Crystallised as the HCl salt from ethanoland diethyl ether.

EXAMEPLE 4 (2R)-2-((R)-[2,6-dichlorophenyl][phenylthio]methyl)morpholinehydrochloride

i)(2R)-2-((R)-[2,6-dichlorophenyl][phenylthio]methyl)-4-benzylmorpholine

Reacted (2R)-2-[(S)-bromo(phenyl)methyl]-4-benzylmorpholine (200 mg,0.58 mmol) (see example 8(iv) ), 2,6-dichlorobenzenethiol (130 mg, 0.70mmol) and anhydrous potassium carbonate (97 mg, 0.70 mmol) followingexample 2(iv). The reaction mixture was purified directly by SCXchromatography eluting with ammonia/methanol (ca. 3M) to give theproduct as an oil (230 mg).

ii) (2R)-2-((R)-[2,6-dichlorophenyl][phenylthio]methyl)imorpholinehydrochloride

Stirred(2R)-2-((R)-[2,6-dichlorophenyl][phenylthio]methyl)-4-benzylmorpholine(230 mg, 0.52 mmol) with solid supported Hunig's base (270 mg, 1.04mmol) and α-chloroethyl chloroformate (152 mg, 1.04 mmol) indichloromethane (4 ml) at room temperature under nitrogen for 3 h.Filtered and concentrated in vacuo, dissolved oil in methanol andstirred at room temperature for 1 h. Evaporated to give a colourlesssolid. NMR (MeOH) 7.32-7.12 (8H, m), 4.62 (1H, d), 4.31-4.22 (1H, m),4.16-4.06 (1H, m), 3.91-3.80 (1H, m), 3.2-2.9 (4H, m).

EXAMPLE 5(2R)-2-((R)-[4-methylphenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

i) (2R)-2-[(S)-(4-metlylphenyl)(hydroxy)methyl]-4-betizymorpholin-3-oneand (2S)-2-[(R)-(4-methylphenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one

To a stirred solution of (+/−)-4-benzylmorpholin-3-one (4.06 g, 21.3mmol) in anhydrous THF (25 ml) under nitrogen at −80° C. was addedlithium diisopropylamide (2.0M, 19.5 ml) solution inheptane/THF/ethylbenzene dropwise, whilst maintaining the reactiontemperature below −65° C. The resulting solution was stirred for afurther 30 minutes at −78° C., before being slowly added overapproximately 45 minutes to a solution of 4-methylbenzaldehyde (3.07 g,25.51 mmol) in anhydrous THF (15 ml) under nitrogen at −78° C., whilstagain maintaining the reaction temperature below −75° C. The resultingyellow solution was stirred at −78° C. for 0.5 hour, before beingallowed to warm to room temperature. The reaction mixture was cautiouslyquenched by addition of saturated ammonium chloride solution (50 ml) andthe THF was evaporated in vacuo from the mixture. The resulting cloudyaqueous solution was extracted with dichloromethane, and the organicextracts were combined, washed with brine, dried over magnesiumsulphate, filtered and the dichloromethane evaporated in vacuo to give athick red oil (9.35 g). After purification by flash columnchromatography (eluent: ethyl acetate/hexane 30/70 to 70/30 gradient[v/v]) the colourless oil obtained was triturated with hexane followedby hot cyclohexane to give after successive decanting of supernatant anddrying the product as a colourless solid (2.46 g).

ii) (S)-[4-methylphenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(R)-[4-methylphenyl][(2R)-4-benzylmorpholin-2-yl]methanol

The product was prepared from(2R)-2-[(S)-(4-methylphenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(4-methylphenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(2.50 g, 8.04 mmol) following the procedure described in EXAMPLE 2(ii).The oil was purified by flash chromatography on silica eluting withethyl acetate/heptane gradient 30/70 to 70/30 to give required productas an oil (1.16 g).

iii) (2R)-2-[(S)-bromo(4-methylphenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(4-methylphenyl)methyl]-4-benzylmorpholine

To a stirred solution of(S)-[4-methylphenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(R)-[4-methylphenyl][(2R)-4-benzylmorpholin-2-yl]methanol (1.16 g, 3.91mmol) and triphenylphosphine (1.54 g, 5.87 mmol) in dry dichloromethanewas added dropwise a solution of carbon tetrabromide (1.95 g, 5.87 mmol)in dichloromethane over a period of 10 min. Further triphenylphosphine(0.5 eq) and carbon tetrabromide (0.5 eq) were added after 0.5 h.Quenched reaction mixture after 2 h with saturated aqueous sodiumbicarbonate. Extracted with dichloromethane, dried extracts overmagnesium sulphate, filtered and evaporated to a red oil. Triturated oilwith diethyl ether, filtered and evaporated to a yellow oily solid. Theoil was purified by flash chromatography on silica eluting with ethylacetate/heptane 20/80 to give the product as an oil (0.61 g)

iv)(2R)-2-((R)-[4-methylphenyl]{[2-rnethoxyphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[4-methylphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine

Reacted (2R)-2-[(S)-bromo(4-methylphenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(4-methylphenyl)methyl]-4-benzylmorpholine (340 mg,0.94 mmol), 2-methoxybenzenethiol (161 mg, 1.13 mmol) and anhydrouspotassium carbonate (160 mg, 1.13 mmol) following procedure described inexample 2(iv). The crude oil was purified by flash chromatography onsilica eluting with ethyl acetate/heptane 20/80 to give the product asan oil (0.21 g).

v) (2R)-2-((R)-(4-methylphenyl){[2-methoxphenyl]thio}methyl)morpholinehydrochloride

Debenzylation of(2R)-2-((R)-[4-methylphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[4-methylphenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine(210 mg, 0.50 mmol) following procedure described in EXAMPLE 1(iv) butat room temperature gave a colourless oil (180 mg) from which the firsteluting enantiomer was obtained after chiral chromatography on aChiralcel-OD column, elutuant heptane/isopropanol/dimethylethylamine(20/80/0.2) Rt 9.70 min. The oil was dissolved in dichloromethane andHCl/diethyl ether added to give the title compound as the hydrochloridesalt (36 mg). NMR (DMSO) 9.20 (2H, br. s), 7.24-7.08 (6H, m), 6.92 (1H,d), 6.89 (1H, t), 4.60 (1H, d), 4.09-3.97 (2H, m), 3.80 (3H, s),3.78-3.66 (1H, m), 3.20-3.13 (1H, m), 3.04-2.90 (3H, m), 2.24 (3H, s).

EXAMPLE 6 (2R)-2-((R)-[phenyl][2-chlorophenylthio]methyl)morpholinehydrochloride

i) (R)-[Phenyl][(2S)-4-benzylmorphoin-2-yl]methyl methanesulphonate and(S)-[Phenyl][(2R)-4-benzylmorpholin-2-yl]methyl methanesulphonate

To a stirred solution of(R)-[phenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(S)-[phenyl][(2R)-4-benzylmorpholin-2-yl]methanol (2.0 g, 7.06 mmol) indry dichloromethane (24 ml) at room temperature under nitrogen was addedtriethylamine (0.78 g, 7.77 mmol) and methanesulphonyl chloride (0.89 g,7.77 mmol). After stirring overnight at room temperature, the reactionmixture was diluted with diethyl ether and filtered. The filtrate wasevaporated to dryness to give an orange oil (2.5 g).

ii) (2R)-2-((R)-[phenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholineand (2S)-2-((S)-[phenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholine

To a stirred suspension of(R)-[phenyl][(2S)-4-benzylmorpholin-2-yl]methyl methanesulphonate and(S)-[phenyl][(2R)-4-benzylmorpholin-2-yl]methyl methanesulphonate (790mg, 2.19 mmol) and anhydrous potassium carbonate (1.50 g, 10.95 mmol) indry degassed DMF (3 ml) under nitrogen at room temperature was added2-chlorobenzenethiol (1.58 g, 10.95 mmol). After stirring at roomtemperature for 18 h, diluted with water and extracted withdichloromethane (2×). The extracts were washed with 2N NaOH, water andbrine, dried over magnesium sulphate, filtered and evaporated to an oil.After purification by flash column chromatography (eluent: heptane/ethylacetate 100/0 to 70/30 [v/v]) the product was obtained as a colourlessoil (0.26 g)

iii) (2R)-2-((R)-[phenyl][2-chlorophenylthio]methyl)morpholinehydrochloride

Debenzylation of(2R)-2-((R)-[phenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholine and(2S)-2-((S)-[phenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholine(250 mg, 0.61 mmol) following procedure described in EXAMPLE 1(iv) butat room temperature gave a colourless oil (190 mg) from which the firsteluting enantiomer was obtained after chiral chromatography on aChiralPak-AD column eluant heptane/ethanoldimethylethylamine (85/15/0.2)Rt 7.55 min. NMR (DMSO) 9.34 (2H, br. s), 7.31-7.00 (9H, m), 4.68 (1H,d), 4.09-3.90 (1H, m), 3.98-3.87 (1H, m), 3.69-3.58 (1H, m), 3.10-3.01(1H, m), 2.90-2.79 (3H, m). Converted to the title product hydrochloridesalt.

EXAMPLE 7 (2R)-2-((R)-[phenyl][2-methylphenylthio]methyl)morpholinehydrochloride

i) (2R)-2-((R)-[phenyl]{[2-methylphenyl]thio}methyl)-4-benzylmorpholineand (2S)-2-((S)-[phenyl]{[2-methylphenyl]thio}methyl)-4-benzylmorpholine

The product was prepared as an oil (0.22 g) from(R)-[phenyl][(2S)-4-benzylmorpholin-2-yl]methyl methanesulphonate and(S)-[phenyl][(2R)-4-benzylmorpholin-2-yl]methyl methanesulphonate (0.49g, 1.46 mmol), 2-methylbenzenethiol (0.22 g, 1.75 mmol) and potassiumcarbonate (0.24 g, 1.75 mmol) following the procedure described inEXAMPLE 6(ii)

ii) (2R)-2-((R)-[phenyl][2-methlylphenylthio]methyl)morpholinehydrochloride

Debenzylation of(2R)-2-((R)-[phenyl]{[2-methylphenyl]thio}methyl)-4-benzylmorpholine and(2S)-2-((S)-[phenyl]{[2-methylphenyl]thio}methyl)-4-benzylmorpholine(210 mg, 0.54 mmol) following procedure described in EXAMPLE 1(iv) butat room temperature gave a colourless oil (180 mg) from which the firsteluting enantiomer was obtained after chiral chromatography on aChiralPak-OJ column eluant heptane/ethanol/dimethylethylamine(40/60/0.2) Rt 8.86 min. This was converted to the title producthydrochloride salt and crystallised from isopropanol/methanol. NMR(CDCl₃) 10.06 (2H, br. s), 7.20-6.93 (9H, m), 4.35-4.27 (1H, m),4.10-3.93 (3H, m), 3.22-3.11 (2H, m), 3.03-2.86 (2H, m), 2.28 (3H, s).

EXAMPLE 8(2R)-2-((R)-[phenyl][2-trifluoromethylphenylthio]methyl)morpholinehydrochloride

Method 1(i) 4-Benzylmorpholin-3-one

A solution of N-benzyl-N-(2-hydroxyethyl)chloroacetamide (1.0 eq., 627.7g, 2.759 mol) in tert-butanol (0.9 L) was stirred under nitrogen whilewarming to 25-30° C. A 1.0 M solution of potassium tert-butoxide intert-butanol (1.05 eq., 2.897 L, 2.897 mol) was added over 2 hours,maintaining the reaction temperature between 30 and 32° C. The reactionmixture was stirred at 27-28° C. for 90 minutes. When TLC showed thereaction to be complete, ice-cold water (6 L) was added and theresultant cloudy solution extracted with EtOAc (1×3 L, 2×1.5 L). Thecombined organic layers were washed with brine (2×3 L), dried over MgSO4and evaporated in vacuo to give a light brown oil (441 g, 84% yield),which was used in the next stage without further purification; MW191.23; C11H13NO2; Rf 0.52 (80% EtOAc, 20% hexane); 1H NMR (CDCl3):7.40-7.29 (5H, m), 4.67 (2H, s), 4.28 (2H, s), 3.87 (2H, t, 5.4 Hz),3.31 (2H, t, 5.4 Hz); LCMS: m/z 192 [M+H]+@ Rt 1.00 min (single majorpeak).(ii) (2R)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]morpholin-3-one,(2S)-4-benzyl-2-[(R)-hydroxy(phenyl)methyl]morpholin-3-one and(2R)-4-benzyl-2-[(R)-hydroxy(phenzyl)methyl]morpholin-3-one,(2S)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]morpholin-3-one

To a stirred solution of 4-benzylmorpholin-3-one (5.02 g, 26 mmol) inanhydrous THF (25 ml) under nitrogen at −78° C. was added a 2 M.solution of LDA in heptane/THF/ethylbenzene (1.5 eq., 39 mmol, 19.5 ml)over approximately 20 minutes, whilst maintaining the reactiontemperature below −75° C. The resulting brown solution was stirred for afurther 30 minutes at −78° C., before being slowly added overapproximately 30 minutes to a solution of benzaldehyde (1.2 eq., 3.34 g,31 mmol) in anhydrous THF (15 ml) under nitrogen at −78° C., whilstagain maintaining the reaction temperature below −75° C. The resultingyellow solution was stirred at −78° C. for 1 hour, before being allowedto warm to room temperature slowly over 1 hour. The reaction mixture wascautiously quenched by addition of saturated ammonium chloride solution(50 ml) and the THF was evaporated in vacuo from the mixture. Theresulting cloudy aqueous solution was extracted with DCM (3×50 ml), andthe organic extracts were combined, washed with brine (50 ml), driedover Na₂SO₄ and the DCM evaporated in vacuo to give a thick brown oil(9.2 g), which partially crystallised on standing. The mixture ofdiastereoisomeric alcohols was purified and separated by flash columnchromatography using gradient elution (from 10% EtOAc, 90% DCM to 20%EtOAc, 80% DCM), which gave(2R)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]morpholin-3-one and(2S)-4-benzyl-2-[(R)-hydroxy(phenyl)methyl]morpholin-3-one as light redcrystals (2.461 g, 31% yield); MW 297.36; C18H19NO3; Rf 0.40 (50% EtOAc,50% hexane); 1H NMR (CDCl3): 7.41-7.36 (2H, m), 7.31-7.16 (6H, m),6.91-6.86 (2H, m), 5.14 (1H, d, J 3.5 Hz), 4.71 (1H, d, 14.5 Hz), 4.48(1H, d, J 3.5 Hz), 4.25 (1H, d, 14.5 Hz), 4.20 (1H, br. s), 3.89 (1H,ddd, 11.7 Hz, 2.5 Hz, 2.0 Hz), 3.67 (1H, dt, 11.2 Hz, 3.4 Hz), 3.16 (1H,dt, 12.0 Hz, 4.0 Hz), 2.86 (1H, br. d, 12.0 Hz); LCMS: m/z 298 [M+H]+ @Rt 1.24 min (single major peak). This reaction was performed on scalesfrom 200 mg to 5 g (yield range 20 to 40%).(2R)-4-benzyl-2-[(R)-hydroxy(phenyl)methyl]morpholin-3-one and(2S)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]morpholin-3-onediastereoisomer was isolated as a brown solid (1.42 g) contaminated withN-benzylmorpholin-3-one. Trituration with EtOAc afforded the purecompound as a white solid (0.484 g, 6% yield); MW 297.36; C18H19NO3; Rf0.23 (50% EtOAc, 50% hexane); 1H NMR (CDCl3): 7.61-7.55 (2H, m),7.50-7.36 (6H, m), 7.31-7.25 (2H, m), 5.21 (1H, d, 2.3 Hz), 5.09 (1H, d,J 7.7 Hz, 2.3 Hz), 4.73 (2H, s, s Hz), 4.37 (1H, d, J 7.7 Hz), 4.01(1H,ddd, 12.0 Hz, 2.6 Hz, 1.9 Hz), 3.77 (1H, dt, 11.0 Hz, 3.5 Hz), 3.50 (1H,dt, 12.0 Hz, 4.0 Hz), 3.16 (1H, br. d, 12.0 Hz); LCMS: m/z 298 [M+H]+ @Rt 1.24 min (single major peak).

(iii) (S)-[(2S)-4-benzylmorpholinyl](phenyl)methanol and(R)-[(2R)-4-benizylmorpholinyl](phenyl)methanol

To a solution of(2R)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]morpholin-3-one and(2S)-4-benzyl-2-[(R)-hydroxy(phenyl)methyl]morpholin-3-one (326 mg, 1.1mmol) in anhydrous THF (5 ml) under nitrogen at room temperature wasslowly added a 1 M solution of borane in THF (4 eq., 4.4 ml, 4.4 mmol).The solution was stirred at 60° C. for 2 hours. After cooling down toroom temperature, dry methanol (2 ml) was slowly added to quench excessborane reagent. 1 M. Aqueous hydrochloric acid solution (2 ml) was addedand the reaction mixture was heated to 60° C. for 1 hour. The organicsolvents were evaporated in vacuo and the concentrated solution waspoured onto 1 M aqueous potassium carbonate solution (10 ml) andextracted with diethyl ether (2×20 ml). The combined organic layers werewashed with brine (20 ml), water (20 ml), dried over MgSO4 andconcentrated in vacuo. The resultant oil was purified by flash columnchromatography (90% hexane, 9% EtOAc, 1% NEt3) to give a viscous oil(189 mg, 60% yield); MW 283.37; C18H21NO2; Rf 0.42 (90% EtOAc, 10%hexane); 1H NMR (CDCl3): 7.45-7.32 (10H, m), 4.67 (1H, d, 7.3 Hz), 4.03(1H, dt, 11.4 Hz, 2.7 Hz), 3.86-3.73 (2H, m), 3.64 (1H, d, 13.2 Hz),3.39 (1H, d, 13.2 Hz), 3.30 (1H, br. s), 2.68 (1H, d, 12.7 Hz), 2.56(1H, d, 10.9 Hz), 2.28-2.15 (2H, m); LCMS: m/z 284 [M+H]+ @ Rt 0.95 min(single major peak).

This reaction was performed on scales from 50 mg to 1.5 g (yieldrange=50 to 84%).

(iv) (R)-[(2S)-4-benzylmorpholinyl](phenyl)methanol and(S)-[(2R)-4-benzylmorpholinyl](phenyl)methanol

Using the procedure described in Example 8(iii) starting from(2R)-4-benzyl-2-[(R)-hydroxy(phenyl)methyl]-morpholin-3-one and(2S)-4-benzyl-2-[(S)-hydroxy(phenyl)methyl]-morpholin-3-one (135 mg,0.51 mmol) the reaction and subsequent purification yielded a viscousoil (98 mg, 68% yield); MW 283.37; C₁₈H₂₁NO₂; Rf 0.52 (100% EtOAc); 1HNMR (CDCl3): 7.28-7.17 (10H, m), 4.80 (1H, d, 4.0 Hz), 3.88 (1H, dt,11.4 Hz, 3.0 Hz), 3.72 (1H, m), 3.68-3.61 (1H, m), 3.50 (1H, d, 13 Hz),3.25 (1H, d, 13 Hz), 2.52 (2H, br. t, 12.0 Hz), 2.17 (1H, t, 11 Hz),2.08 (1H, td, 11 Hz, 3.0 Hz), OH not observed; LCMS: m/z 284 [M+H]+ @ Rt0.98 min (single major peak). This reaction was performed on scales from100 to 400 mg (yield range=60 to 93%).

(v) (2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholine and(2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine

To a solution of (S)-[(2S)-4-benzylmorpholinyl](phenyl)methanol and(R)-[(2R)-4-benzylmorpholinyl](phenyl)methanol (10.27 g, 36.29 mmol) inanhydrous dichloromethane (150 ml) under nitrogen at room temperaturewas added freshly recrystallised triphenylphosphine (1.4 eq., 13.310 g,50.80 mmol) followed by carbon tetrabromide (1.4 eq., 16.849 g, 50.80mmol) as a solution in anhydrous dichloromethane (50 ml). After 15minutes the reaction mixture was diluted with dichloromethane (100 ml)and washed with saturated aqueous sodium hydrogencarbonate solution (150ml), brine (150 ml), dried over MgSO4 and concentrated in vacuo to givean orange oil (42.0 g). To the orange oil was added diethyl ether (200ml) and the resulting suspension was sonicated for 30 minutes. Thesolvent was decanted and the process repeated with a further portion ofdiethyl ether (200 ml). The combined ethereal extracts were concentratedin vacuo to yield an orange solid (22.0 g) which was purified by flashcolumn chromatography (10% EtOAc: 89.5% Hexane, 0.5% Triethylamine) togive a white solid (7.20 g, 58% yield). Alternative Work-up: Thereaction mixture was poured onto a silica (160 g) filtration pad whichwas washed using suction with dichloromethane (14×250 ml). Strippingthis filtrate in vacuo gave crude product (16.0 g, 131% uncorrected).This was purified by flash column chromatography (5% EtOAc: 94.5%Hexane: 0.5% Triethylamine to 10% EtOAc: 89.5% Hexane: 0.5%Triethylamine) to give a white solid (6.05 g, 50% yield); MW 346.27;C18H20BrNO; Rf 0.76 (70% EtOAc, 30% hexane); 1H NMR (CDCl3): 7.39-7.14(10H, m), 4.83 (1H, d, 7.4 Hz), 4.01 (1H, br. t, 8.3 Hz), 3.73 (1H, br.d, 11.1 Hz), 3.60-3.48 (2H, m), 3.39 (1H, d, 12 Hz), 3.20 (1H, d, 11.4Hz), 2.50 (1H, d, 10.4 Hz), 2.07 (2H, t, 10.9 Hz); LCMS: m/z 348/346[M+H]+ @ Rt 1.20 min (single major peak). This reaction was performed onscales from 100 to 400 mg (yield range=60 to 93%).

A sample of racemic (2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholineand (2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine (6.02 g) wasseparated by preparative chiral chromatography (Chiralcel-AD 1 kgcolumn, ethanol:dimethylethylamine 100: 0.3) to give the first elutingenantiomer Rt 23.4 min as an off-white solid (2.89 g) of(2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholine and the secondeluting enantiomer Rt 28.9 min as an off-white solid (2.89 g) of(2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine (2.21 g)

vi)(2R)-2-((R)-[phenyl]{[2-trifluoromethylphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[phenyl]{[2-trifluoromethylphenyl]thio}metlyl)-4-benzylmorpholine

To a stirred solution of 2-trifluoromethylthiophenol (2.469 g, 13.86mmol) and (2R)-2[(S)-bromo(4-methylphenyl)methyl)-4-benzylmorpholine and(2S)-2[(R)-bromo(4-methylphenyl)methyl)-4-benzylmorpholine (4.0 g, 11.55mmol) in anhydrous DMF (60 ml) at room temperature under nitrogen wasadded cesium carbonate (4.14 g, 12.71 g). The reaction mixture washeated at 95° C. for 1 h. The reaction mixture was cooled to roomtemperature, diluted with ethyl acetate, then washed sequentially withwater, brine, dried over magnesium sulphate, filtered and evaporated toa brown oil. The oil was purified by flash column chromatography(eluent: hexane/ethyl acetate gradient 100 to 90/10 [v/v]) to give ayellow oil (4.83 g, 94% yield); MW 444; C₂₅H₂₄F₃NOS; ¹H NMR (CDCl₃):7.60 (1H, dd, 7.2 Hz, 1.4 Hz), 7.17-7.39 (13H, m), 4.50 (1H, d, 7.2 Hz),3.97-4.12 (2H, m), 3.73 (1H, dt, 9.7 Hz, 2.3 Hz), 3.59 (1H, d, 12.6 Hz),3.37 (1H, d, 12.6 Hz), 2.57-2.68 (2H, m); 2.18-2.38 (2H, m); LCMS (2.5minutes method): m/z 445 [M+H]+ @ Rt 1.50 min.

vii) (2R)-2-((R)-[phenyl][2-trifluoromethylphenylthio]methyl)morpholinehydrochloride

The title compound was obtained from(2R)-2-((R)-[phenyl]{[2-trifluoromethylphenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[phenyl]{[2-trifluoromethylphenyl]thio}methyl)-4-benzylmorpholine(5.25 g, 11.84 mmol), solid supported Hunig's base (Argonaut, 3.56mmol/g, 6.64 g, 23.67 mmol, 2 eq.) and α-chloroethyl chloroformate (3.83ml, 35.51 mmol, 3 eq.) in anhydrous dichloromethane (75 ml) at 40° C.following the method described in example 1(iv). After evaporation ofthe methanol solution a light brown solid (5.60 g) was obtained whichwas recrystallised from iso-propanol to give the hydrochloride salt asfine white needles. The hydrochloride salt was suspended in ethylacetate and washed with an aqueous solution of sodium hydroxide (50 mlof a 1M solution). The organic layer was washed with brine, dried overMgSO₄ and concentrated in vacuo to yield the free amine as a colourlessoil (3.10 g, 74% yield); MW 353.41; C₁₈H₁₈F₃NOS; ¹H NMR (CDCl₃): 7.46(1H, d, 7.7 Hz), 7.24 (1H, d, 7.3 Hz), 7.05-7.2 (7H, m), 4.28 (1H, d,7.7 Hz), 3.92 (1H, d, 11.4 Hz), 3.80 (1H, q, 7.0 Hz), 3.58 (1H, dt, 1.82Hz, 11.4 Hz), 2.69-2.87 (2H, m), 2.59 (2H, d, 6.0 Hz), 2.13-1.90 (1H, brs); LCMS (10 minutes method): m/z 354 [M+H]+ @ Rt 5.26 min.

A sample of the racemic free base (1.384 g) was separated by preparativechiral chromatography (Chiralpak-OJ, heptane: isopropanol:dimethylethylamine 70: 30: 0.2) to give the first eluting enantiomer Rt9.5 min (0.57 g) as an oil. Redissolved in diethyl ether (20 ml) andtreated with ethereal hydrogen chloride (2M 0.8 ml) to give a whitesolid (566 mg, mp 240-1° C.) of the title product(2R)-2-((R)-[phenyl][2-trifluoromethylphenylthio]methyl)morpholinehydrochloride.

The second eluting enantiomer Rt 15.8 min was obtained as an oil (0.55g) and similarly converted to the hydrochloride salt(2S)-2-((S)-[phenyl][2-trifluoromethylphenylthio]methyl)morpholinehydrochloride (556 mg mp 244-5° C.). A sample (20 mg) was crystallisedfrom isopropanol (2 ml) allowing the solvent to evaporate slowly overseveral weeks. The crystals were analysed by xray crystallography toconfirm the absolute stereochemistry as (S,S) for the second elutingenantiomer, data is listed in tables 1-6 herein.

Method 2

(i) 4-benzyl-morpholine-2-carbonitrile

A one-liter reactor with mechanical stirring, cooled by ice bath, wascharged with N-benzylethanolamine (172.2 g, 1.14 mol).2-Chloroacrylonitrile (100 g, 1.14 mol) was added dropwise over a periodof 2 minutes. The temperature was kept between 23° C. and 29° C. usingan ice bath, progressively replaced by a water bath at 15° C. Afterstirring at room temperature over night the mixture was dissolved in THFand transferred into a 21 reactor cooled to −5° C. by an ice/NaCl bath.The total volume of THF equalled 1.351. Potassium tert-butoxide (148 g,1.1 eq.) was added in portions over 1 hour, while maintaining thetemperature at 0±2° C. After 1 hour stirring at 0° C. the mixture wasquenched by saturated NaHCO₃ (500 ml). The aqueous layer was extractedwith diethyl ether. The organic layers were dried over MgSO₄ andevaporated to dryness. After percolation of the 250 g dry residue on 1kg SiO₂ (eluent: ethyl acetate/n-heptane gradient 5/95 to 80/100 [v/v])4-benzyl-morpholine-2-carbonitrile was obtained as a clear oil (149.8 g,65%).

(ii) (2S)-(4-Benzyl-morpholin-2-yl)-phenyl-methanone and(2R)-(4-Benzyl-morpholin-2-yl)-phenyl-methanone

A 31 double jacket reactor was charged with4-benzyl-morpholine-2-carbonitrile (135.05 g; 1 eq) and dry diethylether (1.41). Alternatively, toluene may be used in place of diethylether. When Tj=0° C. and Tm=1° C. (Tj=temperature of the jacket,Tm=temperature of the mass), phenyl magnesium chloride (2M sol. in THF,360 ml, 1.08 equiv) was added dropwise over 1 hour. Tm rose to 4° C. andcame back to 2° C. at the end of the addition. Tm was progressivelyraised to 17.5° C. within 45 minutes and the mixture stirred at thistemperature for another 45 minutes. The reactor was cooled down to Tm=2°C. and Tj=0° C. (75 minutes) and hydrochloric acid (700 ml of 5Nsolution) was added in two portions. Tm rose to 33° C. After someminutes, the hydrochloride salt of the ketone crystallised. WhenTm=Tj=room temperature, the triphasic suspension was filtered. Theorganic layer of the mother liquors, which contains impurities, waseliminated. The filtration cake was then washed with methylene chloride(700 ml). This liquor was charged in the reactor with the acid aqueouslayer. Treatment of the hydrochloride salt: After drying under vacuum,164.4 g of the hydrochloride contaminated with MgCl₂ were suspended in abiphasic mixture of water/methylenechloride (500 ml/800 ml). Thesuspension was basified with aqueous sodium hydroxide (75 ml of a 30%solution) under ice bath cooling. Mg(OH)₂ precipitated and the aqueouslayer was extracted with methylene chloride. The organic layers arefiltered on a bed of Celite 512 after adding some Celite to the layersthemselves. The filtered organic phase was dried over MgSO₄ andevaporated to dryness. The ketone crystallizes readily on standing(132.4 g; 70%). Treatment of the mother liquors: The combined organicphases were washed with aqueous sodium hydroxide (750 ml of a 2Nsolution). Celite 512 (160 g) was added to the suspension which was thenfiltrated through bed of Celite. The aqueous layer was separated andextracted with methylene chloride. The combined organic phases weredried over MgSO₄ and evaporated to dryness to provide 35.8 g of productenriched with unreacted nitrile. This fraction could be further purifiedby percolation on SiO₂. (2S)-(4-benzyl-morpholin-2-yl)-phenyl-methanoneand (2R)-(4-benzyl-morpholin-2-yl)-phenyl-methanone were separated usingpreparative chiral chromatography. Alternatively, the two enantiomersmay be separated by fractional crystallization from acetonitrile usingfrom 0.55 to 1 equivalent of dibenzoyltartaric acid to generatediastereoisomeric salts of the title compound. The crystals may becollected by filtration and neutralized with 30% NaOH to afford theoptically enriched title compound.

The title compound may also be prepared using the following one-potsynthesis. A 1600 L GL reactor under N₂ was successively loaded with2-chloroacrylonitrile (33.2 kg, 379 moles) and toluene (114 L) at 21° C.Then, N-benzylethanolamine (57 kg, 377 moles) was added and the reactionmixture was post-agitated at room temperature for about 17 h. Then, themixture was diluted with toluene (336 L), cooled down to −12.4° C. andpotassium t-butoxide (42.3 kg, 377 moles) was added in portions (10)maintaining −13.7° C.≦Tmass≦−2.8° C. The mixture was post-agitated atabout 0° C. for 2.5 h, quenched by adding ultra pure water (142.5 L)maintaining 2.1° C.≦Tmass≦8.7° C. The aqueous layer (176 kg) wasseparated after 35 minutes of post-stirring allowing the mixture toreach 15° C. and the toluene layer was washed with ultra pure water(142.5 L) and the aqueous layer (162 kg) was separated. The organiclayer was then concentrated under reduced pressure (150 mbars)maintaining Tmass≦60° C. in order to distill 162 kg of toluene. Thefiltrates were then diluted with toluene (114 L) and treated with SiO₂(Merck silica gel 60, 0.063-0.1 mm, 74.1 kg) under agitation at roomtemperature for 1.25 h. SiO₂ was filtered and rinsed with toluene (2×114L). Then, the filtrates were concentrated under reduced pressure (150mbars) maintaining Tmass≦60° C. in order to distill 351.8 kg of toluene(KF: 0.01% w/w H₂O).

The solution of 4-Benzyl-morpholine-2-carbonitrile (169.2 kg) wasdiluted with toluene (157 L) and was cooled to 0° C. andphenylmagnesiumchloride (25 wt. % solution in THF, 213 kg, 389 moles,1.36 molar equiv.) was slowly added (over 3.5 h) to the reactionmixture, maintaining the temperature at −3° C.≦Tmass≦7° C. The reactionmixture was post-stirred for 2 hours at Tmass≈0° C. Then, the quench wasperformed by adding acetic acid (8.55 L, Tmass=5→17.2° C.), poststirring 10 minutes and cooling to 5° C. before adding an aceticacid/water mixture (229 L, 33/67 v/v). During the quench, addition wasperformed at such a rate that Tmass did not exceed 20° C. (typicalTmass=4.6° C. to 10.4° C.). The mixture was post-agitated overnight atRT and the aqueous layer (285.8 kg) was extracted.

The toluene layer was cooled to 0° C. and a 5 N NaOH aqueous solution(420.1 kg) was slowly added maintaining the temperature at −2.4°C.≦Tmass≦11° C. The reaction mixture was post-stirred for 1 h and theaqueous layer (494.8 kg) was extracted. The toluene layer wasconcentrated under reduced pressure (50 mbars) maintaining Tmass≦60° C.in order to distill 356.2 kg of toluene and isopropanol (180.4 kg) wasadded. The toluene was stripped off under reduced pressure (100 mbars)maintaining Tmass≦60° C. in order to distill 186.4 kg of toluene andisopropanol (135 kg) was added again to the mixture. A last distillationof toluene was performed under reduced pressure (50 mbars) maintainingTmass≦60° C. in order to distill 131 kg of toluene and isopropanol (49.4kg) was finally added to the mixture and the solution was stirred at RTuntil crystallization (17 minutes).

Ultra pure water was added (125.4 L) and the mixture was stirredovernight at RT and cooled down to about 0° C. for 1 hour. Theprecipitate was filtered and rinsed with a cooled water/isopropanol50/50 v/v solution (76.6 kg). The wet precipitate was dried under vacuumat Tjack=35° C. for 96 hours to obtain the title compound as anoff-white powder with 59% overall yield. The title compound may beresolved by the fractional crystallisation process described above.

(iii) (R)-phenyl[(2R)-4-(phenylmethyl)morpholin-2-yl]methanol

To a stirred solution of (+)-DIP chloride (49.6 g, 155 mmol) in dry TBF(150 ml) under nitrogen was added(2R)-(4-benzyl-morpholin-2-yl)-phenyl-methanone (16.54 g, 58.89 mmol) inone portion. The reaction mixture was stirred at room temperature for 18hours. The mixture was evaporated in vacuo and the crude oil taken up inmethanol and absorbed onto 250 g SCX-2 ion exchange resin. After elutionof borane residues with methanol the product was eluted with 2M ammoniain methanol. Removal of solvent in vacuo yielded the product as yellowoil. (11.23 g, 67%); MW 283.37; C₁₈H₂₁NO₂; ¹H NMR (CDCl₃): 7.32-7.45(10H, m), 4.67 (1H, d, 7.3 Hz), 4.03 (1H, dt, 11.4 Hz, 2.7 H), 3.86-3.73(2H, m), 3.64 (1H, d, 13.2 Hz), 3.39 (1H, d, 13.2 Hz), 3.30 (1H, br. s),2.68 (1H, d, 12.7 Hz), 2.56 (1H, d, 10.9 Hz), 2.28-2.15 (2H, m); LCMS:m/z 284 [M+H]+ @ Rt 0.95 min.

(v) (2R)-2-[(S)-bromo(phenyl)methyl]-4-(phenylmethyl)morpholine

To a solution of (R)-phenyl[(2R)-4-(phenylmethyl)morpholin-2-yl]methanol(11.2 g, 39.58 mmol) in anhydrous chloroform (400 ml) under nitrogen wasadded PPh₃Br₂ (33.41 g, 79.15 mmol). The reaction mixture was heated at60° C. overnight. The mixture was allowed to cool to room temperaturethen washed with saturated aqueous sodium carbonate solution, dried overMgSO₄ and concentrated in vacuo. The resulting residue was purified byflash chromatography on silica (eluent: ethyl acetate:isohexane 1:4) togive a pale yellow oil. Trituration with isohexane gave(2R)-2-[(S)-bromo(phenyl)methyl]-4-(phenylmethyl)morpholine as acolourless solid (8.54 g, 62%); MW 346.27; C₁₈H₂₀BrNO; ¹H NMR (CDCl₃):7.14-7.39 (10H, m), 4.83 (1H, d, 7.4 Hz), 4.01 (1H, br. t, 8.3 Hz), 3.73(1H, br. d, 11.1 Hz), 3.60-3.48 (2H, m), 3.39 (1H, d, 12 Hz), 3.20 (1H,d, 11.4 Hz), 2.50 (1H, d, 10.4 Hz), 2.07 (2H, t, 10.9 Hz); LCMS: (6 minmethod) m/z 346 [M]+ @ Rt 2.51 min.

(vi) (2R)-2-[(S)-bromo(phenyl)methyl]-4-(phenylmethyl)morpholine canthen be Converted to the Title Product(2R)-2-((R)-[phenyl][2-trifluoromethylphenylthio]methyl)morpholinehydrochloride using the above Procedure in example 8 Method 1 (v) and(vi).

EXAMPLE 9 (2R)-2-[(R)-[(2-ethylphenyl)thio](phenyl)methyl]morpholine

i)(2S)-2-[(S)-[(2-ethylphenyl)thio](phenyl)methyl]-4-(phenylmethyl)morpholineand(2R)-2-[(R)-[(2-etlylphenyl)thio](phenyl)methyl]-4-(phenylmethyl)morpholine

Compound(2S)-2-[(S)-[(2-ethylphenyl)thio](phenyl)methyl]-4-(phenylmethyl)morpholineand(2R)-2-[(R)-[(2-ethylphenyl)thio](phenyl)methyl]-4-(phenylmethyl)morpholinewas obtained from 2-ethyl-thiophenol (160 mg, 1.16 mmol) and(2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholine and(2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine (200 mg, 0.58 mmol)following a modification of the method described in example 8(vi) inwhich the reaction mixture was heated to 95° C. for 2 hours. Afterpurification by flash column chromatography (eluent: ethylacetate/hexane 9/1 [v/v]) the product was obtained as a white solid (152mg, 65% yield); MW 403.59; C₂₆H₂₉NOS; ¹H NMR (CDCl₃): 6.96-7.40 (14H,m), 4.22 (1H, d, 7.2 Hz), 3.96-4.01 (2H, m), 3.72 (1H, td, 11.1 Hz, 2.2Hz), 3.52 (1H, d, 13.1 Hz), 3.32 (1H, d, 13.1 Hz), 2.68 (2H, q, 7.7 Hz),2.59 (2H, br d, 11.7 Hz), 2.06-2.21 (2H, m), 1.12 (3H, t, 7.2 Hz); LCMS(2.5 minute method) m/z 404 [M+H]+ @ Rt 1.49 min.

ii) (2R)-2-[(R)-[(2-ethylphenyl)thio](phenyl)methyl]morpholinehydrochloride

Reaction of(2S)-2-[(S)-[(2-ethylphenyl)thio](phenyl)methyl]4-(phenylmethyl)morpholineand(2R)-2-[(R)-[(2-ethylphenyl)thio](phenyl)methyl]-4-(phenylmethyl)morpholinefollowing the method in example 1(iv) gave a viscous yellow oil (213.3mg, 86% yield) from which the title product was obtained after chiralseparation on chiral OD semi-preparative column; LC purity=100% (UV254nm)/100% (ELS); MW 313.47; C₁₉H₂₃NOS; ¹H NMR (CDCl₃): 7.17 (1H, d, 7.6Hz), 7.12-7.05 (5H, m), 7.01 (2H, d, 3.8 Hz), 6.87-6.93 (1H, m), 4.07(1H, d, 8.1 Hz), 3.92-3.97 (1H, m), 3.74-3.80 (1H, m), 3.59 (1H, td,11.4 Hz, 3.0 Hz), 2.80 (1H, td, 12.4 Hz, 3.3 Hz), 2.71 (1H, br. d, 12.1Hz), 2.63-2.54 (4H, m), 1.64 (1H, br. s), 1.04 (3H, t, 7.6 Hz); LCMS (10minutes method): m/z 314 [M+H]+ @ Rt 5.92 min.(2R)-2-[(R)-[(2-ethylphenyl)thio](phenyl)methyl]morpholine was convertedinto its hydrochloride salt. MW 349.93; C₁₉H₂₃NOS.HCl; ¹H NMR (CDCl₃):10.10 (2H, br. s), 7.13-7.28 (8H, m), 7.02-7.08 (1H, m), 4.36 (1H, br.s), 4.014.17 (3H, br. m), 3.16-3.31 (2H, br. m), 2.92-3.09 (2H, br. m),2.71 (2H, q, 7.7 Hz), 1.15 (3H, t, 7.2 Hz).

EXAMPLE 10(2R)-2-[(R)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]morpholinehydrochloride

i)(2S)-2-[(S)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholineand(2R)-2-[(R)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholine

Compound(2S)-2-[(S)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholineand(2R)-2-[(R)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholinewas obtained from 2-methoxy thiophenol (74 μl, 0.574 mmol) and(2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholine and(2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine (181 mg, 0.522 mmol)following the method in example 8(vi) in which the reaction was heatedat 95° C. for 2.5 h. After purification by flash column chromatography(eluent: ethyl acetate/hexane gradient 15/85 to 25/75 [v/v]) the productwas obtained as a viscous yellow oil (175 mg, 83% yield); MW 405.56;C₂₅H₂₇NO₂S; ¹H NMR (CDCl₃): 7.01-7.26 (12H, m), 6.58-6.63 (2H, m), 4.39(1H, d, 7.2 Hz), 3.86-3.91 (2H, m), 3.71 (3H, s), 3.56-3.62 (1H, m),3.42 (1H, d, 10.8 Hz); 3.21 (1H, d, 10.8 Hz), 2.46-2.52 (2H, m),2.01-2.11 (2H, m); LCMS (10 minutes method): m/z 406 [M+H]⁺ @ R_(T)6.09min.

ii) (2R)-2-[(R)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]morpholinehydrochloride

Reaction of(2S)-2-[(S)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholineand(2R)-2-[(R)-{[2-(Methyloxy)phenyl]thio}(phenyl)methyl]-4-(phenylmethyl)morpholine(100 mg, 0.25 mmol) following the method in example 1(iv) gave a viscousyellow oil (60 mg, 77% yield) from which the product was obtained afterchiral separation on a Chiralcel OJ semi-preparative column. LCpurity=100%; MW 315.44; C₁₈H₂₁NO₂S; ¹H NMR (CDCl₃): 7.14-7.34 (7H, m),6.74-6.84 (2H, m), 4.50 (1H, d, 8.2 Hz), 4.10 (1H, d, 10.9 Hz),3.85-4.00 (4H, m), 3.74 (1H, dt, 1.4 Hz, 11.3 Hz), 2.82-3.02 (2H, m),2.66-3.02 (3H, m); LCMS (10 minutes method): 1m/z 316 [M+H]⁺ @ R_(t)4.87min. This was converted to its hydrochloride salt.

EXAMPLE 11(2R)-2-[(R)-{[2-(methylthio)phenyl]thio}(phenyl)methyl]morpholinehydrochloride

i)(2R)-2-((R)-[phenyl]{[2-methylthiophenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[phenyl]{[2-methylthiophenyl]thio}methyl)-4-benzylmorpholine

To a solution of (2R)-4-benzyl-2-[(S)-bromo(phenyl)methyl]morpholine and(2S)-4-benzyl-2-[(R)-bromo(phenyl)methyl]morpholine (4.0 g, 11.55 mmol)and 2-methylsulphenyl-thiophenol (1.2 eq, 2.17 g, 13.86 mmol) inanhydrous DMF (35 ml) at room temperature under nitrogen was addedcesium carbonate (1.18 eq., 14.04 g, 13.63 mmol). The mixture was heatedat 50° C. for 1.5 hours, allowed to cool to room temperature, taken upin methanol and treated with SCX-2 (100 g). The SCX-2 was washed withmethanol. The product was obtained as a white solid (4.92 g) after SCXchromatography (eluent: ammonia/methanol 1/1 [v/v]) and removal ofsolvents in vacuo. Purification by flash column chromatography (eluent:ethyl acetate/isohexane gradient 10/90 to 30/70 [v/v]) gave a whitesolid (4.04 g, 86%); MW 421.63; C₂₇H₂₅NOS₂; ¹H NMR (CDCl₃): 7.03-7.15(6H, m), 6.93-6.99 (2H, m), 6.74 (1H, td, 7.3 Hz, 1.5 Hz), 4.31 (1H, d,7.8 Hz), 3.95 (1H, br. d, 12.1 Hz), 3.83 (1H, td, 8.1 Hz, 3.8 Hz), 3.59(1H, td, 11.1 Hz, 2.8 Hz), 2.82 (1H, td, 12.1 Hz, 3.3 Hz), 2.61-2.75(3H, m), 2.35 (3H, s), 1.73 (1H, br. s); LCMS (6 minutes method): m/z422 [M+H]+ @ Rt 3.36 min.

ii) (2R)-2-[(R)-{[2-(methylthio)phenyl]thio}(phenyl)methyl]morpholinehydrochloride

To a suspension of polymer supported Hunig's base (5.02 g) and(2R)-2-((R)-[phenyl]{[2-methylthiophenyl]thio}methyl)-4-benzylmorpholineand(2S)-2-((S)-[phenyl]{[2-methylthiophenyl]thio}methyl)-4-benzylmorpholine(4.02 g, 9.49 mmol) in dry dichloromethane (70 ml) was addedα-chloroethyl chloroformate (2.93 ml, 28.6 mmol, 3 eq.) at roomtemperature and under nitrogen. The mixture was heated at 40° C. for 1.5hours then left to stir at room temperature overnight. The reactionmixture was filtered and concentrated in vacuo to give a pale orangeliquid. This was taken up in methanol (70 ml) and heated at 40° C. for 2hours. A white solid crashed out of the solution which was purified bySCX chromatography (eluent: ammonia/methanol 1/1 [v/v]). Afterevaporation in vacuo the product was obtained as a pale yellow oil (3.13g, 99%); MW 331.50; C₁₈H₂₁NOS₂; ¹H NMR (CDCl₃): 7.03-7.15 (6H, m),6.93-6.99 (2H, m), 6.74 (1H, td, 7.3 Hz, 1.5 Hz), 4.31 (1H, d, 7.8 Hz),3.95 (1H, br. d, 12.1 Hz), 3.83 (1H, td, 8.1 Hz, 3.8 Hz), 3.59 (1H, td,11.1 Hz, 2.8 Hz), 2.82 (1H, td, 12.1 Hz, 3.3 Hz), 2.61-2.75 (3H, m),2.35 (3H, s), 1.73 (1H, br. s). After separation by chiralchromatography the oil was converted into its hydrochloride salt inwhich the pale yellow oil was taken up in isopropanol (˜200 ml) andfiltered. Addition of hydrogen chloride (19 ml of a 1M solution indiethyl ether, 19 mmol) gave a white precipitate to which furtherdiethyl ether (˜50 ml) was added. The solid was isolated by filtration,washed with diethyl ether give the hydrochloride salt of the titleproduct as a white solid (3.03 g); MW 367.96; C₁₈H₂₂ClNOS₂; ¹H NMR(CDCl₃): 9.94 (2H, br. s), 7.06-7.18 (6H, m), 6.94-7.03 (2H, m), 6.78(1H, t, 6.8 Hz), 4.24-4.32 (1H, m), 4.20 (1H, d, 5.8 Hz), 3.89-4.06 (2H,m), 3.18 (2H, br. t, 11.9 Hz), 2.99 (2H, br. s), 2.37 (3H, s); LCMS (10minutes method): m/z 332 [M−C]+ @ Rt 5.07 min.

EXAMPLE 12(2R)-2-((R)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

A solution of lithium diisopropylanide (2M in heptane, 18.2 ml) wasadded dropwise over 20 min to a stirred solution of4-benzyl-morpholin-3-one (5.0 g, 26 mmol) and 4-chlorobenzaldehyde (4.41g, 31.4 mmol) in dried tetrahydrofuran (60 ml) cooled to −70° C. undernitrogen atmosphere. After 1 h at −70° C., the reaction mixture wasquenched with aqueous ammonium chloride (100 ml) and extracted withethyl acetate (100 ml). The extracts were washed with 2M aqueoushydrochloric acid (100 ml), brine solution (100 ml) and dried oversodium sulphate. After filtration, the solution was evaporated and theresidual oil purified by chromatography on silica eluting with ethylacetate:hexane 70:30 then ethyl acetate to give diastereomer 1(2R)-2-[(S)-(4-chlorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(4-chlorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one asa solid (2.64 g) followed by diastereomer 2(2R)-2-[(R)-(4-chlorophenyl)(hydroxy)methyl]4-benzylmorpholin-3-one and(2S)-2-[(S)-(4-chlorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one asan oil (1.46 g). Diastereomer 1 was recrystallised from ethyl acetate(25 ml) n-hexane (100 ml) to give white needles (2.47 g, 29%)

A solution of borane in tetrahydrofuran (1M, 29 ml) was added dropwiseto a stirred solution of diastereomer 1(2R)-2-[(S)-(4-chlorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(4-chlorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(2.40 g 7.24 mmol) in dried tetrahydrofuran (30 ml) at room temperatureunder nitrogen causing effervesence. The solution was heated to 60° C.for 2 h, allowed to cool to room temperature and excess borane quenchedby adding methanol (14 ml) slowly. Aqueous hydrochloric acid (1M, 14 ml)was added, heated to 60° C. for 1 h and then evaporated to a whitesolid. Added saturated aqueous sodium carbonate (50 ml) and diethylether (50 ml) to dissolve the solid and extracted with diethyl ether(2×50ml). The extracts were washed with brine solution, dried, filteredand evaporated to a colourless oil (2.38 g). The oil was purified bychromatography on silica eluting with diethyl ether:hexane 75:25 to give(R)-[4-chlorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[4-chlorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol as acolourless oil (2.08 g)

A solution of carbon tetrabromide (2.82 g, 8.5 mmol) in dichloromethane(3 ml) was added dropwise over 10 min to a stirred solution of(R)-[4-chlorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[4-chlorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol (1.80 g, 5.67mmol) and triphenylphosphine (2.23 g, 8.5 mmol) in dichloromethane (40ml) at room temperature under nitrogen. After 30 min, the reactionsolution was washed with saturated aqueous sodium bicarbonate (50 ml).The dichloromethane layer was dried, filtered and evaporated to a redliquid (8.5 g). Trituration with diethyl ether (20 ml) crystallisedtriphenylphoshine oxide that was then removed by filtration. Thefiltrate was evaporated to a yellow oil and was purified bychromatography on silica eluting with ethyl acetate:hexane 20:80 to givea colourless oil of(2R)-2-[(S)-bromo(4-chlorophenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(4-chlorophenyl)methyl]-4-benzylmorpholine (1.17 g)that crystallised to a pink solid on standing.

Cesium carbonate (667 mg, 2.05 mmol) was added to a stirred solution of(2R)-2-[(S)-bromo(4-chlorophenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(4-chlorophenyl)methyl]-4-benzylmorpholine (651 mg,1.71 mmol) and 2-methoxybenzenethiol (287 mg, 2.05 mmol) in drydimethylformamide (3 ml). The suspension was heated to 90° C. for 1 h.The cooled reaction mixture was diluted with iced water, 2M aqueoussodium hydroxide (1 ml) and extracted with diethyl ether (15 ml). Theextracts were washed with brine solution, dried, filtered and evaporatedto a yellow oil (0.89 g). The crude product was purified bychromatography on silica eluting with ethyl acetate:heptane 1:4 to give2(R)-2-((R)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineas a pale yellow oil (619 mg, 82%)

Alpha-chloroethyl chloroformate (0.3 ml, 2.78 mmol) was added to agently stirred suspension of polystyrene supported diisopropylethylamine(Argonaut, 390 mg, 1.39 mmol) and2(R)-2-((R)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine(610 mg, 1.39 mmol) in dichloromethane (8 ml) at room temperature undernitrogen. After 3 h, the suspension was filtered and the filtrateevaporated. The residue was dissolved in methanol (10 ml) and heated to60° C. for 1 h. The solution was evaporated and the solid residuecrystallised from isopropanol (5 ml) diethyl ether (10 ml) to give awhite solid (441 mg, 82%). The racemic hydrochloride salt was convertedto the free base by stirring in dicloromethane (20 ml) and aqueoussodium hydroxide (1M, 20 ml). The dichloromethane layer was separated,dried, filtered and evaporated to a colourless oil (403 mg). Chiralpreparative chromatography (Chiralcel-OD,heptane:ethanol:dimethylethylamine 50:50:0.2) was used to isolate thefirst eluting enantiomer Rt 9.3 min as an oil. This was redissolved indiethylether and treated with ethereal hydrogen chloride to give thetitle product(2R)-2-((R)-[4-chlorophenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride as a solid (159 mg, 36%, mp 238-241° C.), NMR (DMSO) 9.31(2H, br. s), 7.32 (4H, dd), 7.07-7.20 (2H, m), 6.91 (1H, d), 6.76 (1H,t), 4.67 (1H, d), 4.0-4.1 (2H, br d), 3.78 (3H, s), 3.72 (1H, t), 3.15(1H, d), 2.95-3.1 (3H, m) LCMS: m/z 350 [M+H]⁺ @ Rt 3.8 min.

EXAMPLE 13(2R)-2-((R)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride

A solution of lithium diisopropylamide (2M in heptane, 16.9 ml) wasadded dropwise over 15 min to a stirred solution of4-benzyl-morpholin-3-one (5.0 g, 26 mmol) and 3-fluorobenzaldehyde (3.55g, 28.6 mmol) in dried tetrahydrofuran (60 ml) cooled to −70° C. undernitrogen atmosphere. After 1 h at −70° C., the reaction mixture wasquenched with aqueous ammonium chloride (100 ml) and extracted withethyl acetate (100 ml). The extracts were washed with 2M aqueoushydrochloric acid (2×50 ml), brine solution (100 ml) and dried oversodium sulphate. After filtration, the solution was evaporated and theresidual oil purified by chromatography on silica eluting with ethylacetate:hexane 70:30 then ethyl acetate to give diastereomer 1(2R)-2-[(S)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one asa solid (3.8 g) followed by diastereomer 2(2R)-2-[(R)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(S)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one asan oil (2.13 g). Diastereomer 1 was recrystallised from ethyl acetate(25 ml) n-hexane (100 ml) to give white needles (2.62 g, 32%).

A solution of borane in tetrahydrofuran (1M, 30.7 ml) was added dropwiseto a stirred solution of diastereomer 1(2R)-2-[(S)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(R)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(2.42 g 7.68 mmol) in dried tetrahydrofuran (30 ml) at room temperatureunder nitrogen causing effervesence. The solution was heated to 60° C.for 2 h, allowed to cool to room temperature and excess borane quenchedby adding methanol (15 ml) slowly. Aqueous hydrochloric acid (1M, 15 ml)was added, heated to 60° C. for 1 h and then evaporated to a whitesolid. Added saturated aqueous sodium carbonate (50 ml) and diethylether (50 ml) to dissolve the solid and extracted with diethyl ether(2×50 ml). The extracts were washed with brine solution, dried, filteredand evaporated to a colourless oil (2.38 g). The oil was purified bychromatography on silica eluting with diethyl ether:hexane 75:25 to give(R)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol as acolourless oil (2.23 g)

A solution of carbon tetrabromide (3.3 g, 9.96 mmol) in dichloromethane(4 ml) was added dropwise over 5 min to a stirred solution of(R)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol (2.0 g, 6.64mmol) and triphenylphosphine (2.61 g, 9.96 mmol) in dichloromethane (40ml) at room temperature under nitrogen. After 30 min, the reactionsolution was washed with saturated aqueous sodium bicarbonate (50 ml).The dichloromethane layer was dried, filtered and evaporated to a redliquid (8.5 g). Trituration with diethyl ether (40 ml) crystallisedtriphenylphoshine oxide that was then removed by filtration. Thefiltrate was evaporated to a yellow oil and was purified bychromatography on silica eluting with ethyl acetate:hexane 20:80 to givea colourless oil of(2R)-2-[(S)-bromo(3-fluorophenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(3-fluorophenyl)methyl]-4-benzylmorpholine (0.811 g,34%).

Cesium carbonate (546 mg, 1.68 mmol) was added to a stirred solution of(2R)-2-[(S)-bromo(3-fluorophenyl)methyl]-4-benzylmorpholine and(2S)-2-[(R)-bromo(3-fluorophenyl)methyl]-4-benzylmorpholine (510 mg, 1.4mmol) and 2-methoxybenzenethiol (235 mg, 1.68 mmol) in drydimethylformamide (3 ml). The suspension was stirred at room temperaturefor 4 h. The reaction mixture was diluted with iced water, 2M aqueoussodium hydroxide (1 ml) and extracted with diethyl ether (15 ml). Theextracts were washed with brine solution, dried, filtered and evaporatedto a yellow oil (627 mg). The crude product was purified bychromatography on silica eluting with ethyl acetate:heptane 20:80 togive2(R)-2-((R)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineas a pale yellow oil (466 mg, 79%)

Alpha-chloroethyl chloroformate (0.235 ml, 2.17 mmol) was added to agently stirred suspension of polystyrene supported diisopropylethylamine(Argonaut, 306 mg, 1.09 mmol) and2(R)-2-((R)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)-4-benzylmorpholine(460 mg, 1.09 mmol) in dichloromethane (6 ml) at room temperature undernitrogen. After 16 h, the suspension was filtered and the filtrateevaporated. The residue was dissolved in methanol (6 ml) and heated to60° C. for 1 h. The solution was evaporated and the solid residuecrystallised from isopropanol (15 ml) and n-hexane to give a white solid(371 mg, 92%). The racemic hydrochloride salt was converted to the freebase by stirring in dicloromethane (20 ml) and aqueous sodium hydroxide(0.5M, 20 ml). The dichloromethane layer was separated, dried, filteredand evaporated to a colourless oil (341 mg). Chiral preparativechromatography (Chiralcel-OD, heptane:ethanol:dimethylethylamine50:50:0.2) was used to isolate the first eluting enantiomer Rt 9.2 minas an oil. This was redissolved in diethylether and treated withethereal hydrogen chloride to give the title product(2R)-2-((R)-[3-fluorophenyl]{[2-methoxyphenyl]thio}methyl)morpholinehydrochloride as a solid (138 mg, 34%, mp 233-234° C.). NMR (DMSO) 9.26(2H, br. s), 7.31 (1H, q), 7.10-7.20 (4H, m), 7.05 (1H, t), 6.92 (1H,d), 6.78 (1H, t), 4.66 (1H, d), 4.0-4.15 (2H, m), 3.77 (3H, s), 3.72(1H, t), 3.19 (1H, d), 2.92-3.1 (3H, m). LCMS: m/z 334 [M+H]⁺ @ Rt 3.5min

EXAMPLE 14(2R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)morpholinehydrochloride

A solution of borane in tetrahydrofuran (1M, 25.2 ml) was added dropwiseto a stirred solution of diastereomer 2 from example 13(i)(2R)-2-[(R)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and(2S)-2-[(S)-(3-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(2.0 g 6.3 mmol) in dried tetrahydrofuran (25 ml) at room temperatureunder nitrogen causing effervesence. The solution was heated to 60° C.for 1.5 h, allowed to cool to room temperature and excess boranequenched by adding methanol (10 ml) slowly. Aqueous hydrochloric acid(1M, 13 ml) was added, heated to 60° C. for 1 h and then evaporated to awhite solid. Added saturated aqueous sodium carbonate (50 ml) anddiethyl ether (50 ml) to dissolve the solid and extracted with diethylether (2×50 ml). The extracts were washed with brine solution, dried,filtered and evaporated to a colourless oil (2.01 g). The oil wasdissolved in isopropanol (20 ml) and ethereal hydrogen chloride (2M, 3ml) added to crystallise the salt(R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol hydrochlorideand (S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanolhydrochloride as a white solid (1.66 g, 78%)

Methanesulphonyl chloride (1.01 g, 8.9 mmol) was added dropwise over 5min to a stirred solution of(R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol hydrochlorideand (S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanolhydrochloride (1.50 g, 4.44 mmol) and triethylamine (1.79 g, 17.8 mmol)in dry dichloromethane (30 ml) at room temperature under nitrogenatmosphere. After 1 h, water (30 ml) was added, stirred vigorously andthen the dichloromethane layer separated. The solution was dried oversodium sulphate, filtered and evaporated to a colourless oil. The oilwas purified by chromatography on silica eluting with diethylether:hexane 3:1 to give(R)-[3-fluorophenyl][(2S)4-benzylmorpholin-2-yl]methyl methanesulphonateand (S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate as a colourless oil (1.51 g, 90%)

Anhydrous potassium carbonate (276 mg, 2 mmol) was added to a stirredsolution of (R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methylmethanesulphonate and(S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate (682 mg, 1.8 mmol) and 2-ethoxybenzenethiol (308 mg, 2mmol) in dry dimethylformamide (13 ml) at room temperature undernitrogen atmosphere. After 3 days, water (25 ml) was added and themixture extracted with diethyl ether (25 ml). The extracts were washedwith brine solution (20 ml), dried, filtered and evaporated to a yellowoil (0.89 g). The product was purified by chromatography on silicaeluting with ethyl acetate:heptane 20:80 to give2(R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholineas a colourless oil (493 mg, 63%).

Debenzylation of2(R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholine(480 mg, 1.1 mmol) using the method described in example 13(v) gave theracemic hydrochloride salt (360 mg, 85%). After conversion to the freebase, chiral preparative chromatography (Chiralcel-OD,heptane:ethanol:dimethylethylamine 80:20:0.2) was used to isolate thefirst eluting enantiomer Rt 14.9 min as an oil. This was redissolved indiethylether and treated with ethereal hydrogen chloride to give thetitle product(2R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)morpholinehydrochloride as a solid (144 mg, 34%, mp 211-215° C.). NMR (DMSO) 9.30(2H, br. s), 7.31 (1H, q), 7.11-7.20 (4H, m), 7.05 (1H, t), 6.91 (1H,d), 6.79 (1H, t), 4.64 (1H, d), 3.97-4.15 (4H, m), 3.72 (1H, t), 3.19(1H, d), 2.92-3.1 (3H, m), 1.37 (3H, t). LCMS: m/z 348 [M+H]⁺ @ Rt 4.1min

A solution of 3-fluorophenylmagnesium bromide (0.5M, 50 ml, 25 mmol) wasadded dropwise to a stirred solution of 4-benzyl-mopholin-2-carbonitrile(4.59 g, 22.7 mmol) in diethyl ether (50 ml) at 0° C. under nitrogen.After 45 min at 0 C. the mixture was allowed to warm to room temperaturefor 30 min then recooled and quenched by the addition of aqueoushydrochloric acid (5M, 40 ml)—caution exothermic. After 30 min at roomtemperature, the acidic mixture was basified with sodium hydroxide (5M,60 ml) and extracted with ethyl acetate (3×150 ml). The combinedextracts were washed with brine solution, dried, filtered andconcentrated in vacuo to give(+/−)-(4-benzyl-morpholin-2-yl)-(3-fluorophenyl)methanone (6.9 g) as ayellow oil.

Borontrifluoride etherate (27.6 g, 194 mmol) followed by trifluoroaceticacid (40 ml) were added to a stirred solution of(+/−)-(4-benzyl-morpholin-2-yl)-(3-fluorophenyl)methanone (23.28 g, 77mmol) and triphenylsilane (81.1 g, 311 mmol) in dichloromethane (1000ml) at 0° C. under nitrogen. After 16 h at room temperature, reactionmixture was cooled and carefully basified by addition of aqueous sodiumbicarbonate. The organic layer was separated, dried and concentrated invacuo. The residual orange oil was purified using SCX-2 resin to absorbthe amine product. Elution with methanolic ammonia (2M) andconcentration in vacuo gave an oil (30 g) that was further purified bychromatography on silica (toluene:diethyl ether 60:40) to give(R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol and(S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol as an oil(19.7 g, 84%).

Convertion of (R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanoland (S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol (19.7 g)using method 1 in example 14(ii) gave(R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methylmethanesulphonate and(S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate (24.7 g) was followed by method 1 in example 14(iii)to give2(R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)-4-benzylmorpholine(22.5 g). This was debenzylated by method 1 in example 14(iv) to give(2R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)morpholine and(2S)-2-((S)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)morpholine(15.2 g) followed by preparative chiral chromatography to separate thefirst eluting enantiomer (7.7 g) and then salt formation to the titleproduct(2R)-2-((R)-[3-fluorophenyl]{[2-ethoxyphenyl]thio}methyl)morpholinehydrochloride (4.38 g) as a white crystalline solid.

EXAMPLE 15(2R)-2-((R)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)morpholinehydrochloride

Anhydrous potassium carbonate (377 mg, 2.73 mmol) was added to a stirredsolution of (R)-[3-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methylmethanesulphonate and(S)-[3-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methylmethanesulphonate (740 mg, 1.8 mmol) and 2-chlorobenzenethiol (393 mg,2.73 mmol) in dry dimethylformamide (10 ml) at room temperature undernitrogen atmosphere. After 44 h, diluted with methanol (15 ml) and theinorganic solid filtered. The filtrate was poured directly onto SCX-2columns (3×10 g), washed with methanol and the basic product eluted withmethanolic ammonia (2M) to give a yellow oil (707 mg) after evaporation.The product was further purified by chromatography on silica elutingwith ethyl acetate:heptane 20:80 to give2(R)-2-((R)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholineas a colourless oil (653 mg, 78%).

Alpha-chloroethyl chloroformate (0.33 ml, 3.06 mmol) was added to agently stirred suspension of polystyrene supported diisopropylethylamine(Argonaut, 429 mg, 1.53 mmol) and2(R)-2-((R)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholineand2(S)-2-((S)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)-4-benzylmorpholine(429 mg, 1.53 mmol) in dichloromethane (10 ml) at room temperature undernitrogen. After 2 h, the suspension was filtered and the filtrateevaporated. The residue was dissolved in methanol (10 ml) and heated to60° C. for 1 h. The solution was evaporated to the hydrochloride salt,redissolved in methanol and converted to the free base using SCX-2column (10 g) eluting with methanol and then methanolic ammonia (2M) togive a colourless oil (501 mg, 97%). Chiral preparative chromatography(Chiralcel-OJ, heptane:isopropanol:dimethylethylamine 90:10:0.2) wasused to isolate the first eluting enantiomer Rt 19.2 min as an oil. Thiswas redissolved in diethylether and treated with ethereal hydrogenchloride to give the title product(2R)-2-((R)-[3-fluorophenyl]{[2-chlorophenyl]thio}methyl)morpholinehydrochloride as a solid (231 mg, 40%, mp 183-7° C.). NMR (DMSO) 9.38(2H, br. s), 7.15-7.47 (7H, m), 7.09 (1H, t), 4.85 (1H, d), 4.12-4.20(1H, m), 4.08 (1H, d), 3.77 (1H, t), 3.20 (1H, d), 2.95-3.10 (3H, m).LCMS: m/z 338/340 [M+H⁺ @ Rt 3.9 min

EXAMPLE 16(2R)-2-[(R)-(2-chloro-6-methylphenyl)thio](phenyl)methyl]morpholinehydrochloride

To a solution of (2R)-4-benzyl-2[(S)-bromo(phenyl)methyl]morpholine (200mg, 0.6 mmol) and 2-chloro-6-methyl thiophenol (0.167 ml, 6 eq) inanhydrous DMF (5 ml) at room temperature under nitrogen was addedpotassium carbonate (100 mg, 0.7 mmol, 1.2 eq). The reaction mixture wasstirred at room temperature for 5 hours. The reaction mixture wasdiluted with methanol and poured directly onto a SCX-2 column forpurification to give(2R)-2-[(R)-(2-chloro-6-methylphenyl)thio](phenyl)methyl]-4-(phenyl)methyl]morpholinebefore taking directly onto the next step.

ii) (2R)-2-[(R)-(2-chloro-6 methylphenyl)thio](phenyl)methyl]morpholinehydrochloride

To a suspension of polymer supported Hunig's base (182 mg, 3 eq) and(2R)-2-[(R)-(2-chloro-6methylphenyl)thio](phenyl)methyl]-4-(phenyl)methyl]morpholine (254 mg,0.6 mmol) in dry DCM (5 ml) was added α-chloroethyl chloroformate (0.187ml, 1.7 mmol, 3 eq) at room temperature and under nitrogen. The mixturewas allowed to stir at room temperature overnight. The reaction mixturewas taken up in methanol (5 ml) and stirred at room temperatureovernight. The reaction mixture was then treated with SCX-2 (10 g).After elution with methanol followed by elution with 7 N NH₃/methanol(2R)-2-[(R)-(2-chloro-6 methylphenyl)thio](phenyl)methyl]morpholine wasobtained as an oil (163 mg, 82% yield); MW 333; C₁₈H₂₀ClNOS; ¹H NMR(DMSO): 8.80 (1H, br s), 7.30 (1H, m), 7.20 (7H, m), 4.40 (1H, d, 8.2Hz), 4.20 (1H, m), 4.00 (1H, m), 3.80 (1H, m), 3.15 (1H, m), 2.90 (2H,m), 2.20 (3H, s) 1.20 (1H, m); LCMS (10 minutes method): m/z 334[M+H]⁺ @R_(T)5.1 min; HPLC purity=100% (UV_(215nm))/100% (ELS). The free basewas converted into the title product HCl salt.

EXAMPLE 17(2R)-2-((R)-[4-fluorophenyl]{2-methoxyphenyl]thio}methyl)morpholinehydrochloride

Following the procedure described in example 5(i),4-benzylmorpholin-3-one (4.06 g) was converted to2-(R)-2-[(S)-(4-fluorphenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one and2-(S)-2-[(R)-(4-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one.Crystallised from hexane-ethyl acetate to give a colourless solid (2.04g).

2-(R)-2-[(S)-(4-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-oneand2-(S)-2-[(R)-(4-fluorophenyl)(hydroxy)methyl]-4-benzylmorpholin-3-one(2.0 g) was converted to(R)-[4-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[4-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol followingprocedure described in example 2(ii) to give a colourless oil (1.88 g).

To a stirred solution of(R)-[4-fluorophenyl][(2R)-4-benzylmorpholin-2-yl]methanol and(S)-[4-fluorophenyl][(2S)-4-benzylmorpholin-2-yl]methanol (1.64 g, 5.54mmol)) and triphenylphosphine (2.32 g, 8.86 mmol) in anhydrouschloroform (40 ml) was added solid carbon tetrabromide (2.76 g, 8.31mmol) in one lot. The solution was heated at reflux under nitrogen for 3h. Cooled and washed reaction mixture with brine, dried, filtered andevaporated to a red oil. The oil was purified by chromatography onsilica eluting with hexane:ethyl acetate 41:9 to give2(R)-2-[(S)-bromo(4-fluorophenyl)methyl]-4-benzylmorpholine and2(S)-2-[(R)-bromo(4-fluorophenyl)methyl]-4-benzylmorpholine as acolourless oil (0.49 g)

To a stirred suspension of2(R)-2-[(S)-bromo(4-fluorophenyl)methyl]-4-benzylmorpholine and2(S)-2-[(R)-bromo(4-fluorophenyl)methyl]-4-benzylmorpholine (0.6 g, 1.65mmol) and cesium carbonate (0.59 g, 1.81 mmol) in dry DMF (5 ml) wasadded 2-methoxybenzenethiol (0.25 g, 1.81 mmol). The suspension washeated at 90° C. under nitrogen for 3 h. The cooled reaction mixture wasdiluted with water and extracted with diethyl ether. The extracts werewashed with water and brine, dried, filtered and evaporated to an oil.The crude oil was purified by chromatography on silica eluting withhexane:ethyl acetate 4:1 then 3:2 to give(2R)-2-((R)-(4-fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholine and(2S)-2-((S)-(4-fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholine acolourless oil (0.22 g)

Reaction of the mixture of(2R)-2-((R)-(4-fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholine and(2S)-2-((S)-(4-fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholine(430 mg, 1.02 mmol) following procedure described in EXAMPLE 1(iv) gavea colourless oil (340 mg, 90% yield) from which first eluting enantiomer(2R)-2-((R)-(4-fluorophenyl){[2-methoxyphenyl]thio}methyl)morpholine wasobtained after chiral chromatography on a Chiralcel-OD column eluantheptane/isopropanol/dimethylethylamine (50/50/0.2): Rt 10.58 min. Thiswas converted into its hydrochloride salt. ¹H NMR (CD₃OD): 7.01-7.20(4H, m), 6.70-6.80 (3H, m), 6.60 (1H, t), 4.37 (1H, d,), 3.82-3.90 (1H,m), 3.70-3.79 (4H, m), 3.49-3.60 (1H, m), 2.70-2.78 (2H, m),.2.60-2.70(2H, m).

The pharmacological profile of the present compounds may be demonstratedas follows. All of the exemplified compounds above have been found toexhibit a K_(i) value less than 100 nM at the serotonin transporter anda K_(i) value less than 100 nM at the norepinephrine transporter asdetermined using the scintillation proximity assays described below.Furthermore, all of the exemplified compounds above have been found toselectively inhibit the serotonin and norepinephrine transportersrelative to the dopamine transporter by a factor of at least five usingthe scintillation proximity assays as described below.

Generation of Stable Cell-lines Expressing the Human Dopamine,Norepinephrine and Serotonin Transporters

Standard molecular cloning techniques were used to generate stablecell-lines expressing the human dopamine, norepinephrine and serotonintransporters. The polymerase chain reaction (PCR) was used in order toisolate and amplify each of the three full-length cDNAs from anappropriate cDNA library. Primers for PCR were designed using thefollowing published sequence data:

Human dopamine transporter: GenBank M95167. Reference: Vandenbergh D J,Persico A M and Uhl G R. A human dopamine transporter cDNA predictsreduced glycosylation, displays a novel repetitive element and providesracially-dimorphic TaqI RFLPs. Molecular Brain Research (1992) volume15, pages 161-166.

Human norepinephrine transporter: GenBank M65105. Reference: PacholczykT, Blakely, R D and Amara S G. Expression cloning of a cocaine- andantidepressant-sensitive human noradrenaline transporter. Nature (1991)volume 350, pages 350-354.

Human serotonin transporter: GenBank L05568. Reference: Ramamoorthy S,Bauman A L, Moore K R, Han H, Yang-Feng T, Chang A S, Ganapathy V andBlakely R D. Antidepressant- and cocaine-sensitive human serotonintransporter: Molecular cloning, expression, and chromosomallocalization. Proceedings of the National Academy of Sciences of the USA(1993) volume 90, pages 2542-2546.

The PCR products were cloned into a mammalian expression vector (egpcDNA3.1 (Invitrogen)) using standard ligation techniques. Theconstructs were then used to stably transfect HEK293 cells using acommercially available lipofection reagent (Lipofectamine™—Invitrogen)following the manufacture's protocol.

Scintillation Proximity Assays for Determining the Affinity of TestLigands at the Norepinephrine and Serotonin Transporters.

The compounds of the present invention are norepinephrine and serotoninreuptake inhibitors, and possess excellent activity in, for example, ascintillation proximity assay (e.g. J. Gobel, D. L. Saussy and A. Goetz,J. Pharmacol. Toxicolo. (1999), 42, 237-244). Thus ³H-nisoxetine bindingto norepinephrine re-uptake sites in a cell line transfected with DNAencoding human norepinephrine transporter binding protein and similarly³H-citalopram binding to serotonin re-uptake sites in a cell linetransfected with DNA encoding human serotonin transporter bindingprotein have been used to determine the affinity of ligands at thenorepinephrine and serotonin transporters respectively.

Norepinephrine Binding Assay

Membrane Preparation:

Cell pastes from large scale production of HEK-293 cells expressingcloned human norepinephrine transporters were homogenized in 4 volumes50 mM Tris-HCl containing 300 mM NaCl and 5 mM KCl, pH 7.4. Thehomogenate was centrifuged twice (40,000 g, 10 min, 4° C.) with pelletre-suspension in 4 volumes of Tris-HCl buffer containing the abovereagents after the first spin and 8 volumes after the second spin. Thesuspended homogenate was centrifuged (100 g, 10 min, 4° C.) and thesupernatant kept and re-centrifuged (40,000 g, 20 min, 4° C.). Thepellet was resuspended in Tris-HCl buffer containing the above reagentsalong with 10% w/v sucrose and 0.1 mM phenylmethylsulfonyl fluoride(PMSF). The membrane preparation was stored in aliquots (1 ml) at −80°C. until required. The protein concentration of the membrane preparationwas determined using a bicinchoninic acid (BCA) protein assay reagentkit (available from Pierce).

[³H]-Nisoxetine Binding Assay:

Each well of a 96 well microtitre plate was set up to contain thefollowing:

-   50 μl 2 nM [N-methyl-³H]-Nisoxetine hydrochloride (70-87 Ci/mmol,    from NEN Life Science Products)-   75 μl Assay buffer (50 mM Tris-HCl pH 7.4 containing 300 mM NaCl and    5 mM KCl)-   25 μl Test compound, assay buffer (total binding) or 10 μm    Desipramine HCl (non-specific binding)-   50 μl Wheatgerm agglutinin coated poly(vinyltoluene) (WGA PVT) SPA    Beads (Amersham Biosciences RPNQ0001) (10 mg/ml)-   50 μl Membrane (0.2 mg protein per ml)

The microtitre plates were incubated at room temperature for 10 hoursprior to reading in a Trilux scintillation counter. The results wereanalysed using an automatic spline fitting programmed (Multicalc,Packard, Milton Keynes, UK) to provide Ki values for each of the testcompounds.

Serotonin Binding Assay

The ability of a test compound to compete with [³H]-citalopram for itsbinding sites on cloned human serotonin transporter containing membraneshas been used as a measure of test compound ability to block serotoninuptake via its specific transporter (Ramamoorthy, S., Giovanetti, E.,Qian, Y., Blakely, R., (1998) J. Biol. Chem. 273, 2458).

Membrane Preparation:

Membrane preparation is essentially similar to that for thenorepinephrine transporter containing membranes as described above. Themembrane preparation was stored in aliquots (1 ml) at −70° C. untilrequired. The protein concentration of the membrane preparation wasdetermined using a BCA protein assay reagent kit.

[³H]-Citalopram Binding Assay:

Each well of a 96 well microtitre plate was set up to contain thefollowing:

-   50 μl 2 nM [³H]-Citalopram (60-86 Ci/mmol, Amersham Biosciences)-   75 μl Assay buffer (50 mM Tris-HCl pH 7.4 containing 150 mM NaCl and    5 mM KCl)-   25 μl Diluted compound, assay buffer (total binding) or 100 μM    Fluoxetine (non-specific binding)-   50 μl WGA PVT SPA Beads (40 mg/ml)-   50 μl Membrane preparation (0.4 mg protein per ml)

The microtitre plates were incubated at room temperature for 10 hoursprior to reading in a Trilux scintillation counter. The results wereanalysed using an automatic spline fitting programme (Multicalc,Packard, Milton Keynes, UK) to provide Ki (nM) values for each of theunknown compounds.

Dopamine Binding Assay

The ability of a test compound to compete with [³H]-WIN35,428 for itsbinding sites on human cell membranes containing cloned human dopaminetransporter has been used as a measure of the ability of such testcompounds to block dopamine uptake via its specific transporter(Ramamoorthy et al 1998 supra).

Membrane Preparation:

Is essentially the same as for membranes containing cloned humanserotonin transporter as described above.

[³H]-WIN35,428 Binding Assay:

Each well of a 96well microtitre plate was set up to contain thefollowing:

-   50 μl 4 nM [³H]-WIN35,428 (84-87 Ci/mmol, from NEN Life Science    Products)-   75 μl Assay buffer (50 mM Tris-HCl pH 7.4 containing 150 mM NaCl and    5 mM KCl)-   25 μl Diluted compound, assay buffer (total binding) or 100 μM    Nomifensine (non-specific binding)-   50 μl WGA PVT SPA Beads (10 mg/ml)-   50 μl Membrane preparation (0.2 mg protein per ml.)

The microtitre plates were incubated at room temperature for 120 minutesprior to reading in a Trilux scintillation counter. The results wereanalysed using an automatic spline fitting programme (Multicalc,Packard, Milton Keynes, UK) to provide Ki values for each of the unknowncompounds.

Formalin Paw Assay

The analgesic effect of compounds of the invention for the treatment ofpersistent nociceptive pain was demonstrated using the well-known“formalin test.” The formalin test is a model of persistent nociceptiveactivation induced by tissue injury which can lead to centralsensitization. (Shibata, M., Ohkubo, T., Takahashi, H., and Inoki, R.,“Modified formalin test: Characteristic biphasic pain response,” Pain(1989) 38: 347-352; and Tjolsen, A., Berge, O. G., Hunskaar, S.,Rosland, J. H., and Hole, K., “The formalin test: an evaluation of themethod,” Pain (1992) 51:5-17. ) The effect of compounds of the inventionon formalin-induced paw-licking behavior in the rat was investigated asan index of persistent nociceptive activation. In this test, theinjection of formalin under the skin on the dorsal lateral surface ofthe hind paw of rats causes an immediate and intense increase in thespontaneous activity of C fiber afferents. This activation evokes adistinctly quantifiable behavior indicative of pain, such as licking ofthe injected paw. The behavioral response to formalin is biphasic, withan early phase that is short lived, followed by an extended tonicresponse or late phase of persistent nociceptive activation. Mechanismscausing the late phase response, such as central sensitization of paintransmitting neurons, are currently believed to contribute to varioustypes of persistent pains.

Male Sprague-Dawley rats (200-250 g; Charles River, Portage, Mich.) weremaintained at constant temperature and light (12 h light/12 h dark) for4-7 days prior to the studies. Animals had free access to food and waterat all times prior to the day of the experiment.

The formalin test was performed in custom made Plexiglas® boxes 25×25×20cm (length×width×height) in size. A mirror placed at the back of the boxallowed the unhindered observation of the formalin injected paw. Ratswere acclimatized individually in the cubicles at least 1 hour prior tothe experiment. All testing was conducted between 08:00 and 14:00 hr andthe testing room temperature was maintained at 21-23° C. Test compoundwas administered 30 or 60 minutes prior to the formalin injection.Formalin (50 μl of a 5% solution in saline) was injected subcutaneouslyinto the dorsal lateral surface of the right hind paw with a 27 gaugeneedle. Observation started immediately after the formalin injection.Formalin-induced pain was quantified by recording in 5 minute intervalsthe number of formalin injected paw licking events and the number ofseconds each licking event lasted. These recordings were made for 50minutes after the formalin injection. Scoring in the formalin test wasperformed according to Coderre et al., 1993b and Abbott et al., 1995.(Coderre T. J., Fundytus M. E., McKenna J. E., Dalal S. and Melzack R.“The formalin test: a validation of the weighted-scores method of thebehavioral pain rating,” Pain(1993b) 54: 43-50; and Abbott F. V.,Franklin K. B. J. and Westbrook R. F. “The formalin test: scoringproperties of the first and second phases of the pain response in rats,”Pain (1995) 60: 91-102.) The sum of time spent licking in seconds fromtime 0 to 5 minutes was considered the early phase while the late phasewas taken as the sum of seconds spent licking from 15 to 40 minutes.

In Vitro Determination of the Interaction of Compounds with CYP2D6 inHuman Hepatic Microsomes

Cytochrome P450 2D6 (CYP2D6) is a mammalian enzyme which is commonlyassociated with the metabolism of around 30% of pharmaceuticalcompounds. Moreover, this enzyme shows a genetic polymorphism with as aconsequence a presence in the population of poor and normalmetabolizers. A low involvement of CYP2D6 in the metabolism of compounds(i.e. the compound being a poor substrate of CYP2D6) is desirable inorder to reduce any variability from subject to subject in thepharmacokinetics of the compound. Also, compounds with a low inhihibitorpotential for CYP2D6 are desirable in order to avoid drug-druginteractions with co-administered drugs that are substrates of CYP2D6.Compounds may be tested both as substrates and as inhibitors of thisenzyme by means of the following assays.

CYP2D6 Substrate Assay

Principle:

This assay determines the extent of the CYP2D6 enzyme involvement in thetotal oxidative metabolism of a compound in microsomes. Preferredcompounds of the present invention exhibit less than 75% totalmetabolism via the CYP2D6 pathway.

For this in vitro assay, the extent of oxidative metabolism in humanliver microsomes (HLM) is determined after a 30 minute incubation in theabsence and presence of Quinidine, a specific chemical inhibitor ofCYP2D6. The difference in the extent of metabolism in absence andpresence of the inhibitor indicates the involvement of CYP2D6 in themetabolism of the compound.

Materials and Methods:

Human liver microsomes (mixture of 20 different donors, mixed gender)were acquired from Human Biologics (Scottsdale, Ariz., USA). Quinidineand β-NADPH (β-Nicotinamide Adenine Dinucleotide Phosphate, reducedform, tetrasodium salt) were purchased from Sigma (St Louis, Mo., USA).All the other reagents and solvents were of analytical grade. A stocksolution of the new chemical entity (NCE) was prepared in a mixture ofAcetonitrile/Water to reach a final concentration of acetonitrile in theincubation below 0.5%.

The microsomal incubation mixture (total volume 0.1 mL) contained theNCE (4 μM), β-NADPH (1 mM), microsomal proteins (0.5 mg/mL), andQuinidine (0 or 2 μM) in 100 mM sodium phosphate buffer pH 7.4. Themixture was incubated for 30 minutes at 37° C. in a shaking waterbath.The reaction was terminated by the addition of acetonitrile (75 μL). Thesamples were vortexed and the denaturated proteins were removed bycentrifugation. The amount of NCE in the supernatant was analyzed byliquid chromatography/mass spectrometry (LC/MS) after addition of aninternal standard. A sample was also taken at the start of theincubation (t=0), and analysed similarly.

Analysis of the NCE was performed by liquid chromatography/massspectrometry. Ten μL of diluted samples (20 fold dilution in the mobilephase) were injected onto a Spherisorb CN Column, 5 μM and 2.1 mm×100 mm(Waters corp. Milford, Mass., USA). The mobile phase consisting of amixture of Solvent A/Solvent B, 30/70 (v/v) was pumped (Alliance 2795,Waters corp. Milford, Mass., USA) through the column at a flow rate of0.2 ml/minute. Solvent A and Solvent B were a mixture of ammoniumformate 5.10⁻³ M pH 4.5/methanol in the proportions 95/5 (v/v) and 10/90(v/v), for solvent A and solvent B, respectively. The NCE and theinternal standard were quantified by monitoring their molecular ionusing a mass spectrometer ZMD or ZQ (Waters-Micromass corp, Machester,UK) operated in a positive electrospray ionisation.

The extent of CYP2D6 involvement (% of CYP2D6 involvement) wascalculated comparing the extent of metabolism in absence and in presenceof quinidine in the incubation.

The extent of metabolism without inhibitor (%) was calculated asfollows:

$\frac{\begin{matrix}{{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 0} -} \\{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 30}\end{matrix}}{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 0} \times 100$

The extent of metabolism with inhibitor (%) was calculated as follows:

$\frac{\begin{matrix}{{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 0} -} \\{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 30}\end{matrix}}{\left( {{NCE}\mspace{14mu}{response}\mspace{14mu}{in}\mspace{14mu}{samples}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right){time}\mspace{14mu} 0} \times 100$where the NCE response is the area of the NCE divided by the area of theinternal standard in the LC/MS analysis chromatogram, time0 and time30correspond to the 0 and 30 minutes incubation time.

The % of CYP2D6 involvement was calculated as follows:

$\frac{\begin{matrix}{\left( {\%\mspace{14mu}{extent}\mspace{14mu}{of}\mspace{14mu}{metabolism}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right) -} \\\left( {\%\mspace{14mu}{extent}\mspace{14mu}{of}\mspace{14mu}{metabolism}\mspace{14mu}{with}\mspace{14mu}{inhibitor}} \right)\end{matrix}}{\%\mspace{14mu}{extent}\mspace{14mu}{of}\mspace{14mu}{metabolism}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \times 100$CYP2D6 Inhibitor AssayPrinciple:

The CYP2D6 inhibitor assay evaluates the potential for a compound toinhibit CYP2D6. This is performed by the measurement of the inhibitionof the bufuralol 1′-hydroxylase activity by the compound compared to acontrol. The 1′-hydroxylation of bufuralol is a metabolic reactionspecific to CYP2D6. Preferred compounds of the present invention exhibitan IC₅₀ higher than 6 μM for CYP2D6 activity, the IC₅₀ being theconcentration of the compound that gives 50% of inhibition of the CYP2D6activity.

Material and methods:

Human liver microsomes (mixture of 20 different donors, mixed gender)were acquired from Human Biologics (Scottsdale, Ariz.). β-NADPH waspurchased from Sigma (St Louis, Mo.). Bufuralol was purchased fromUltrafine (Manchester, UK). All the other reagents and solvents were ofanalytical grade.

Microsomal incubation mixture (total volume 0.1 mL) contained bufuralol10 μM, β-NADPH (2 mM), microsomal proteins (0.5 mg/mL), and the newchemical entity (NCE) (0, 5, and 25 μM) in 100 mM sodium phosphatebuffer pH 7.4. The mixture was incubated in a shaking waterbath at 37°C. for 5 minutes. The reaction was terminated by the addition ofmethanol (75 μL). The samples were vortexed and the denaturated proteinswere removed by centrifugation. The supernatant was analyzed by liquidchromatography connected to a fluorescence detector. The formation ofthe 1′-hydroxybufuralol was monitored in control samples (0 μM NCE) andin the samples incubated in presence of the NCE. The stock solution ofNCE was prepared in a mixture of Acetonitrile/Water to reach a finalconcentration of acetonitrile in the incubation below 1.0%.

The determination of 1′ hydroxybufuralol in the samples was performed byliquid chromatograhy with fluorimetric detection as described below.Twenty five μL samples were injected onto a Chromolith PerformanceRP-18e column (100 mm×4.6 mm) (Merck KGAa, Darmstadt, Germany). Themobile phase, consisting of a mixture of solvent A and solvent B whosethe proportions changed according the following linear gradient, waspumped through the column at a flow rate of 1 ml/min:

Time (minutes) Solvent A (%) Solvent B (%) 0 65 35 2.0 65 35 2.5 0 1005.5 0 100 6.0 65 35

Solvent A and Solvent B consisted of a mixture of 0.02 M potassiumdihydrogenophosphate buffer pH3/methanol in the proportion 90/10 (v/v)for solvent A and 10/90 (v/v) for solvent B. The run time was 7.5minutes. Formation of 1′-hydroxybufuralol was monitored by fluorimetricdetection with extinction at λ 252 nm and emission at λ 302 nm.

The IC₅₀ of the NCE for CYP2D6 was calculated by the measurement of thepercent of inhibition of the formation of the 1′-hydroxybufuralol inpresence of the NCE compared to control samples (no NCE) at a knownconcentration of the NCE.

The percent of inhibition of the formation of the 1′-hydroxybufuralol iscalculated as follows:

$\frac{\begin{matrix}{\left( {1^{\prime}\text{-}{hydroxybufuralol}\mspace{14mu}{formed}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right) -} \\\left( {1^{\prime}\text{-}{hydroxybufuralol}\mspace{14mu}{formed}\mspace{14mu}{with}\mspace{14mu}{inhibitor}} \right)\end{matrix}}{\left( {1^{\prime}\text{-}{hydroxybufuralol}\mspace{14mu}{area}\mspace{14mu}{formed}\mspace{14mu}{without}\mspace{14mu}{inhibitor}} \right)} \times 100$

The IC₅₀ is calculated from the percent inhibition of the formation ofthe 1′-hydroxybufuralol as follows (assuming competitive inhibition):

$\frac{{NCE}\mspace{14mu}{Concentration} \times \left( {100 - {{Percent}\mspace{14mu}{of}\mspace{14mu}{inhibition}}} \right)}{{Percent}\mspace{14mu}{of}\mspace{14mu}{inhibition}}$

The IC₅₀ estimation is assumed valid if inhibition is between 20% and80% (Moody G C, Griffin S J, Mather A N, McGinnity D F, Riley R J. 1999.Fully automated analysis of activities catalyzed by the major humanliver cytochrome P450 (CYP) enzymes: assessment of human CYP inhibitionpotential. Xenobiotica, 29(1): 53-75).

X-Ray Crystallographic Data

TABLE 1 Crystal data and structure refinement for 2003xf. Identificationcode 2003xf Empirical formula C18H19ClF3NOS Formula weight 389.85Temperature 107(2) K Wavelength 0.71073 A Crystal system, space groupMonoclinic, P2(1) Unit cell dimensions a = 9.984(2) A alpha = 90 deg. b= 5.6484(13) A beta = 100.867(4) deg. c = 15.931(4) A gamma = 90 deg.Volume 882.4(4) A{circumflex over ( )}3 Z, Calculated density 2, 1.467Mg/m{circumflex over ( )}3 Absorption coefficient 0.371 mm{circumflexover ( )}−1 F(000) 404 Crystal size .06 × .08 × .18 mm Theta range fordata collection 1.30 to 28.20 deg. Limiting indices 11 <= h <= 13, −7 <=k <= 7, −20 <= l <= 19 Reflections collected/unique 5986/3378 [R(int) =0.0661] Completeness to theta = 28.20 92.9% Absorption correction NoneRefinement method Full-matrix least-squares on F{circumflex over ( )}2Data/restraints/parameters 3378/1/234 Goodness-of-fit on F{circumflexover ( )}2 0.846 Final R indices [l > 2sigma(l)] R1 = 0.0488, wR2 =0.0908 R indices (all data) R1 = 0.1227, wR2 = 0.1101 Absolute structureparameter 0.11(10) Largest diff. peak and hole 0.548 and −0.444e.A{circumflex over ( )}−3

X-Ray Crystallographic Data

TABLE 2 Atomic coordinates (×10{circumflex over ( )}4) and equivalentisotropic displacement parameters (A{circumflex over ( )}2 ×10{circumflex over ( )}3) for 2003xf. U(eq) is defined as one third ofthe trace of the orthogonalized Uij tensor. x y z U(eq) S(8) 8641(1)5291(2) 2641(1) 35(1) O(1) 10279(3) 2645(5) 4200(2) 24(1) C(7) 9992(5)3088(8) 2678(3) 25(1) F(3) 5136(4) 4842(7) 443(2) 65(1) N(4) 13055(4)1352(9) 4386(3) 21(1) C(5) 12147(4) 1431(8) 3536(3) 22(1) F(2) 7264(4)4253(5) 644(2) 51(1) C(20) 10490(5) 1794(8) 1263(3) 31(1) F(1) 6497(4)7227(5) 1228(2) 48(1) C(15) 10669(5) 3416(8) 1925(3) 24(1) C(6) 11008(5)3187(8) 3525(3) 24(1) C(16) 11472(5)  5394(10) 1846(3) 32(1) C(10)6184(5) 3389(9) 1805(3) 26(1) C(13) 5978(5)  382(11) 3117(4) 40(1) C(9)7190(5) 3438(9) 2506(3) 30(1) C(3) 12283(5)  976(8) 5085(3) 27(1) C(12)4992(5)  364(10) 2423(3) 31(1) C(2) 11168(5) 2787(9) 5010(3) 28(1) C(21)6263(6)  4934(11) 1033(4) 41(2) C(18) 11846(5)  4080(10) 494(3) 33(1)C(17) 12048(5) 5721(9) 1131(4) 36(1) C(19) 11078(5) 2138(9) 552(4) 35(1)C(11) 5062(5) 1943(9) 1738(4) 42(2) C(14) 7065(6)  1852(10) 3160(4)43(2) Cl(1) 4131(1) 6360(2) 4214(1) 30(1)

X-Ray Crystallographic Data

TABLE 3 Bond lengths [A] and angles [deg] for 2003xf. S(8)—C(9) 1.767(5)S(8)—C(7) 1.828(5) O(1)—C(2) 1.424(5) O(1)—C(6) 1.440(5) C(7)—C(15)1.495(6) C(7)—C(6) 1.528(6) F(3)—C(21) 1.318(6) N(4)—C(5) 1.481(5)N(4)—C(3) 1.484(6) C(5)—C(6) 1.507(6) F(2)—C(21) 1.337(6) C(20)—C(19)1.385(7) C(20)—C(15) 1.383(6) F(1)—C(21) 1.343(6) C(15)—C(16) 1.395(6)C(16)—C(17) 1.382(7) C(10)—C(9) 1.354(6) C(10)—C(11) 1.374(7)C(10)—C(21) 1.520(8) C(13)—C(12) 1.334(6) C(13)—C(14) 1.358(7)C(9)—C(14) 1.397(7) C(3)—C(2) 1.500(6) C(12)—C(11) 1.421(7) C(18)—C(19)1.351(7) C(18)—C(17) 1.360(7) C(9)—S(8)—C(7) 100.6(2) C(2)—O(1)—C(6)110.4(4) C(15)—C(7)—C(6) 112.3(4) C(15)—C(7)—S(8) 109.4(3)C(6)—C(7)—S(8) 111.5(3) C(5)—N(4)—C(3) 112.0(4) N(4)—C(5)—C(6)  11.2(4)C(19)—C(20)—C(15) 121.2(5) C(20)—C(15)—C(16) 117.1(5) C(20)—C(15)—C(7)121.1(5) C(16)—C(15)—C(7) 121.8(5) O(1)—C(6)—C(5) 109.7(4)O(1)—C(6)—C(7) 107.9(4) C(5)—C(6)—C(7) 111.1(4) C(17)—C(16)—C(15)121.2(5) C(9)—C(10)—C(11) 122.9(5) C(9)—C(10)—C(21) 121.0(5)C(11)—C(10)—C(21) 116.0(5) C(12)—C(13)—C(14) 120.3(6) C(10)—C(9)—C(14)116.4(5) C(10)—C(9)—S(8) 125.2(4) C(14)—C(9)—S(8) 118.4(4)N(4)—C(3)—C(2) 109.0(4) C(13)—C(12)—C(11) 119.7(5) O(1)—C(2)—C(3)111.1(4) F(3)—C(21)—F(1) 107.1(5) F(3)—C(21)—F(2) 105.6(5)F(1)—C(21)—F(2) 105.4(5) F(3)—C(21)—C(10) 113.2(5) F(1)—C(21)—C(10)113.6(5) F(2)—C(21)—C(10) 111.4(5) C(19)—C(18)—C(17) 120.6(5)C(18)—C(17)—C(16) 119.8(5) C(18)—C(19)—C(20) 120.2(5) C(10)—C(11)—C(12)118.1(5) C(13)—C(14)—C(9) 122.5(5) Symmetry transformations used togenerate equivalent atoms:

X-Ray Crystallographic Data

TABLE 4 Anisotropic displacement parameters (A{circumflex over ( )}2 ×10{circumflex over ( )}3) for 2003xf. The anisotropic displacementfactor exponent takes the form: −2 pi{circumflex over ( )}2 [h{circumflex over ( )}2 a*{circumflex over ( )}2 U11 + . . . + 2 h k a*b* U12 ] U11 U22 U33 U23 U13 U12 S(8) 24(1) 24(1) 53(1) −1(1) −1(1) 4(1)O(1) 24(2) 23(2) 24(2) 3(2) 0(2) −2(2) C(7) 20(3) 23(2) 27(3) −3(2)−8(3) 0(2) F(3) 55(2) 88(3) 42(2) 15(2) −16(2) −13(2) N(4) 19(2) 14(2)31(3) 3(2) 3(2) −3(3) C(5) 22(3) 16(2) 26(3) −4(2) 2(2) 2(3) F(2) 69(3)53(2) 39(2) 5(2) 29(2) 3(2) C(20) 29(3) 28(3) 31(3) −12(3) −5(3) −1(2)F(1) 61(2) 35(2) 46(2) 5(2) 5(2) 5(2) C(15) 20(3) 22(3) 27(3) 2(3) −3(2)5(2) C(6) 23(3) 17(2) 33(3) −1(2) 11(3) 1(2) C(16) 40(3) 22(2) 31(3)−3(3) 1(3) −7(3) C(10) 20(3) 30(3) 27(3) 2(3) 8(3) 4(3) C(13) 33(3)45(3) 42(4) 3(3) 7(3) 0(3) C(9) 20(3) 38(3) 31(4) −8(3) 2(3) 7(3) C(3)22(3) 28(3) 32(3) 10(2) 5(2) 0(2) C(12) 22(3) 29(2) 41(4) −1(3) 8(3)−7(3) C(2) 28(3) 34(3) 22(3) −2(3) 3(3) 4(2) C(21) 27(4) 50(4) 43(4)−16(3) −1(3) 10(3) C(18) 24(3) 44(3) 30(4) −1(3) 3(3) 11(3) C(17) 42(4)26(3) 40(4) 0(3) 9(3) −6(2) C(19) 33(3) 38(3) 33(4) −9(3) 2(3) 6(3)C(11) 20(3) 49(4) 52(4) −18(3) −3(3) 8(3) C(14) 35(4) 72(5) 22(3) 16(3)−1(3) −4(3) Cl(1) 24(1) 16(1) 46(1) 1(1) −1(1) −1(1)

X-Ray Crystallographic Data

TABLE 5 Hydrogen coordinates (×10{circumflex over ( )}4) and isotropicdisplacement parameters (A{circumflex over ( )}2 × 10{circumflex over( )}3) for 2003xf. x y z U(eq) H(7A)  9558 1486 2630 30 H(5A) 11757 −1623392 26 H(5B) 12685 1877 3099 26 H(20A)  9954  420 1297 37 H(6A) 113984819 3611 29 H(16A) 11626 6536 2292 38 H(13A)  5919 −637 3583 48 H(3A)12902 1128 5645 33 H(3B) 11886 −636 5043 33 H(12A)  4246 −700 2387 37H(2A) 10639 2529 5468 34 H(2B) 11575 4389 5085 34 H(18A) 12248 4302   540 H(17A) 12584 7087 1084 43 H(19A) 10941 1005  103 42 H(11A)  4354 19981248 50 H(14A)  7767 1799 3653 52 H(4B) 13680(60) 2600(100) 4430(30)53(19) H(4A) 13580(50)  230(90) 4400(30) 29(17)

X-Ray Crystallographic Data

TABLE 6 Torsion angles [deg] for 2003xf. C(9)—S(8)—C(7)—C(15) 115.5(4)C(9)—S(8)—C(7)—C(6) −119.7(4) C(3)—N(4)—C(5)—C(6) 52.2(6)C(19)—C(20)—C(15)—C(16) −0.4(7) C(19)—C(20)—C(15)—C(7) 177.8(4)C(6)—C(7)—C(15)—C(20) 126.4(5) S(8)—C(7)—C(15)—C(20) −109.2(4)C(6)—C(7)—C(15)—C(16) −55.5(6) S(8)—C(7)—C(15)—C(16) 68.9(5)C(2)—O(1)—C(6)—C(5) 60.7(5) C(2)—O(1)—C(6)—C(7) −178.1(4)N(4)—C(5)—C(6)—O(1) −55.1(5) N(4)—C(5)—C(6)—C(7) −174.3(4)C(15)—C(7)—C(6)—O(1) −175.0(4) S(8)—C(7)—C(6)—O(1) 61.9(4)C(15)—C(7)—C(6)—C(5) −54.7(5) S(8)—C(7)—C(6)—C(5) −177.8(3)C(20)—C(15)—C(16)—C(17) 0.7(7) C(7)—C(15)—C(16)—C(17) −177.4(5)C(11)—C(10)—C(9)—C(14) 2.6(8) C(21)—C(10)—C(9)—C(14) −176.4(5)C(11)—C(10)—C(9)—S(8) −178.8(4) C(21)—C(10)—C(9)—S(8) 2.2(7)C(7)—S(8)—C(9)—C(10) −114.6(5) C(7)—S(8)—C(9)—C(14) 64.0(5)C(5)—N(4)—C(3)—C(2) −52.6(6) C(14)—C(13)—C(12)—C(11) −1.9(8)C(6)—O(1)—C(2)—C(3) −63.3(5) N(4)—C(3)—C(2)—O(1) 58.2(5)C(9)—C(10)—C(21)—F(3) −173.8(5) C(11)—C(10)—C(21)—F(3) 7.1(7)C(9)—C(10)—C(21)—F(1) −51.3(7) C(11)—C(10)—C(21)—F(1) 129.6(5)C(9)—C(10)—C(21)—F(2) 67.4(7) C(11)—C(10)—C(21)—F(2) −111.6(5)C(19)—C(18)—C(17)—C(16) 0.5(8) C(15)—C(16)—C(17)—C(18) −0.7(8)C(17)—C(18)—C(19)—C(20) −0.2(8) C(15)—C(20)—C(19)—C(18) 0.1(8)C(9)—C(10)—C(11)—C(12) −2.7(8) C(21)—C(10)—C(11)—C(12) 176.3(5)C(13)—C(12)—C(11)—C(10) 2.3(8) C(12)—C(13)—C(14)—C(9) 1.9(8)C(10)—C(9)—C(14)—C(13) −2.1(8) S(8)—C(9)—C(14)—C(13) 179.2(4) Symmetrytransformations used to generate equivalent atoms

1. A compound of formula (I)

wherein R is H; Ar is an aromatic group selected from unsubstitutedphenyl or phenyl substituted with 1, 2, 3, 4 or 5 substitutents selectedfrom C₁-C₄ alkyl, O(C₁-C₄ alkyl), S(C₁-C₄ alkyl), halo, and phenyloptionally substituted with halo, C₁-C₄ alkyl or O(C₁-C₄ alkyl); X isunsubstituted phenyl or phenyl substituted with 1, 2, 3, 4 or 5substituents selected from C₁-C₄ alkyl, O(C₁-C₄ alkyl), and halo; R′ isH or C₁-C₄ alkyl; each R¹ is independently H or C₁-C₄ alkyl; whereineach above-mentioned C₁-C₄ alkyl group is optionally substituted withone or more halo atoms; or a pharmaceutically acceptable salt thereof.2. A compound as claimed in claim 1, represented by formula II:

in which R₂ and R₃ are each independently selected from H, C₁-C₄ alkyl,O(C₁-C₄ alkyl), S(C₁-C₄ alkyl), halo, and phenyl; and R₄ is selectedfrom H, C₁-C₄ alkyl, O(C₁-C₄ alkyl) and halo; wherein eachabove-mentioned C₁-C₄ alkyl group is optionally substituted with one ormore halo atoms; or a pharmaceutically acceptable salt thereof.
 3. Acompound as claimed in claim 2, wherein R₂ is selected from C₁-C₂ alkyl,O(C₁-C₂ alkyl), S(C₁-C₂ alkyl), Cl and F wherein each above-mentionedC₁-C₂ alkyl group is optionally substituted with one or more halo atoms.4. A compound as claimed in claim 2 or claim 3, wherein R₃ is selectedfrom H, Me and Cl.
 5. A compound as claimed in claim 4, wherein R₄ isselected from H, C₁-C₂ alkyl, O(C₁-C₂ alkyl), Cl and F wherein eachabove-mentioned C₁-C₂ alkyl group is optionally substituted with one ormore halo atoms.
 6. A composition comprising a compound as claimed inclaim 1 or 2 together with a pharmaceutically acceptable diluent,excipient or carrier.
 7. A method for treating persistent pain,comprising administering to a patient in need thereof an effectiveamount of a compound as claimed in claim 1.